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  ~ Table of Contents - Current issue
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February 2015
Volume 33 | Issue 5 (Supplement)
Page Nos. 1-173

Online since Friday, February 06, 2015

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EDITORIAL  

Publishing supplement of a Journal p. 1
A Kapil
DOI:10.4103/0255-0857.150857  PMID:25657122
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REVIEW ARTICLE Top

Regulatory categories of probiotics across the globe: A review representing existing and recommended categorization Highly accessed article p. 2
M Arora, A Baldi
DOI:10.4103/0255-0857.150868  PMID:25657150
Probiotics are friendly live microorganisms (in most cases, bacteria) that are similar to beneficial micro-organisms found in the human gut, whenever consumed, have potential to confer benefit to the health of consumers by maintaining, or improving their intestinal microbial flora and are available to consumers mainly in the form of dietary supplements and foods. All-time high interest in the field of probiotics is due to emerging probiotic industry. Probiotics are available in foods and dietary supplements, even as pharmaceutical formulations (capsules, tablets and powders) and in some other forms as well, but their claims of health benefits may challenge the traditional border between food and medicine. A number of probiotic products have been already introduced into the international market as food supplements, dietary supplements, natural health products, functional foods and many more other categories; as a result, the position of regulatory system for probiotics within existing categories become vague and quite unclear. Common terminology for probiotic products has become a necessity to achieve adequate regulatory control for discussion of probiotic-related issues among government, producers and consumers. The lack of a consistent terminology across the globe leads to legal uncertainty and confusion instead of being a direct obstacle for development of a mature market. This article will explain differences in regulatory categorizations across the globe; discuss the terms like food and drugs with a close relationship to probiotics, the problems associated with unsatisfactorily approached categorization as well as suggestive consolidations for the new categorization which will demarcate probiotics into categories explaining their nutritive claims, health claims or both.
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ORIGINAL ARTICLES Top

Design and development of compact monitoring system for disaster remote health centres p. 11
S Santhi, GS Sadasivam
DOI:10.4103/0255-0857.150871  PMID:25657125
Purpose: To enhance speedy communication between the patient and the doctor through newly proposed routing protocol at the mobile node. Materials and Methods: The proposed model is applied for a telemedicine application during disaster recovery management. In this paper, Energy Efficient Link Stability Routing Protocol (EELSRP) has been developed by simulation and real time. This framework is designed for the immediate healing of affected persons in remote areas, especially at the time of the disaster where there is no hospital proximity. In case of disasters, there might be an outbreak of infectious diseases. In such cases, the patient's medical record is also transferred by the field operator from disaster place to the hospital to facilitate the identification of the disease-causing agent and to prescribe the necessary medication. The heterogeneous networking framework provides reliable, energy efficientand speedy communication between the patient and the doctor using the proposed routing protocol at the mobile node. Results: The performance of the simulation and real time versions of the Energy Efficient Link Stability Routing Protocol (EELSRP) protocol has been analyzed. Experimental results prove the efficiency of the real-time version of EESLRP protocol. Conclusion: The packet delivery ratio and throughput of the real time version of EELSRP protocol is increased by 3% and 10%, respectively, when compared to the simulated version of EELSRP. The end-to-end delay and energy consumption are reduced by 10% and 2% in the real time version of EELSRP.
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Comparison of conventional, immunological and molecular techniques for the diagnosis of symptomatic congenital human cytomegalovirus infection in neonates and infants p. 15
A Choudhary, SK Pati, RK Patro, AK Deorari, L Dar
DOI:10.4103/0255-0857.150874  PMID:25657137
Purpose: Human cytomegalovirus (HCMV) is the commonest pathogen causing congenital infection globally. The diagnosis of congenital infection is based either on viral isolation (in cell culture) or demonstration of HCMV DNA from the urine. Saliva is also being used as an alternative sample to urine for the same. The objective of this study was to compare the following assays-polymerase chain reaction (PCR) from urine, saliva and blood, serology (anti-HCMV IgM) and antigen detection (HCMV pp65 antigenaemia) for the diagnosis of congenital HCMV infection. Materials and Methods: Urine and blood samples were collected from 31 infants (median age: 13 weeks) with suspected HCMV infection. For 18 infants, additional saliva samples were collected and all the above assays were compared. Results: PCR for HCMV DNA from urine and anti-HCMV IgM were performed for all 31 infants. Of these, 22 (70.9%) were positive for both assays. In 18 (of the 22) infants positive by both assays, PCR for HCMV DNA from saliva was positive in all 18 (100%), PCR from blood in 7/18 (38.8%) and HCMV pp65 antigenaemia only in 1/18 (5.5%) of the infants. Conclusion: Detection of HCMV DNA in urine combined with anti-HCMV IgM are suitable assays to diagnose HCMV infection in infants. Both PCR from the blood and HCMV pp65 antigenaemia lack sensitivity in infants. Salivary PCR combines convenience with high sensitivity and can substitute PCR from urine, especially in the outpatient and field settings. To the best of our knowledge, this is the first study from India to evaluate salivary PCR for the diagnosis of congenital HCMV infection.
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The predictive value of early indicators for HBeAg seroconversion in HBeAg-positive chronic hepatitis B patients with Telbivudine treatment for 104 weeks p. 20
Juan Wang, Ling-yao Du, Xia Zhu, En-qiang Chen, Hong Tang
DOI:10.4103/0255-0857.148827  PMID:25657151
Purpose: Through an observation on HBeAg-positive chronic hepatits B (CHB) patients in Telbivudine (LDT) treatment for 104 weeks, we tried to explore valuable early predictors for HBeAg seroconversion during the treatment. Materials and Methods: A prospective study lasting for 104 weeks was conducted, and the patients enrolled were administered with LDT 600 mg daily. The medical evaluation went every 12 weeks, then the age distribution, baseline ALT level, early HBVDNA, HBsAg and HBeAg levels at baseline, week 12 and 24 as well as the decrease of the three indicators at week 12 and 24 were analyzed for their predictive values for HBeAg seroconversion at week 104. Result: Thirty-three patients finished the observation. All patients got ALT normalisation and 28 patients (84.84%) got complete virological response (HBV DNA < 291 copies/ml) at week 104. Poor virological response and virologic breakthrough was observed in two (6.06%) and three patients (9.09%), respectively. Nine patients (27.27%) got HBeAg seroconversion. HBeAg levels and its decrease levels at week 12 and 24 showed significant differences between patients with and without HBeAg seroconversion. And the HBsAg levels at week 12 and 24 showed tendencies of significant differences in two groups. HBeAg level at week 24 was confirmed related to its longer term seroconversion in regression analysis. The patients with HBeAg level < 2.1 S/CO at week 24 would be more possible to get HBeAg seroconversion at week 104, with sensitivity, specificity, positive and negative predictive value of 95.83%, 88.89%, 95.8% and 88.9%, respectively. Conclusion: Good efficacy of long-term LDT treatment in biological and virological response and its advantage in serological response was confirmed again in our study. The HBeAg level at week 24 showed significant value in prediction for HBeAg seroconversion at week 104 compared to other serological markers in the early period.
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Performance of rapid influenza diagnostic tests (QuickVue) for Influenza A and B Infection in India  Highly accessed article p. 26
PA Koul, H Mir, MA Bhat, UH Khan, MM Khan, MS Chadha, RB Lal
DOI:10.4103/0255-0857.148831  PMID:25657152
Background: Rapid point-of-care (POC) tests provide an economical alternative for rapid diagnosis and treatment of influenza, especially in public health emergency situations. Objectives: To test the performance of a rapid influenza diagnostic test, QuickVue (Quidel) as a POC test against a real-time polymerase chain reaction (RT-PCR) assay for detection of influenza A and B in a developing country setting. Study Design: In a prospective observational design, 600 patients with influenza-like illness (ILI) or with severe acute respiratory illness (SARI) who were referred to the Influenza Clinic of a tertiary care hospital in Srinagar, India from September 2012 to April 2013, were enrolled for diagnostic testing for influenza using QuickVue or RT-PCR. All influenza A-positive patients by RT-PCR were further subtyped using primers and probes for A/H1pdm09 and A/H3. Results: Of the 600 patients, 186 tested positive for influenza A or B by RT-PCR (90 A/H1N1pdm09, 7 A/H3 and 89 influenza B), whereas only 43 tested positive for influenza (influenza A = 22 and influenza B = 21) by QuickVue. Thus, the sensitivity of the QuickVue was only 23% (95% confidence interval, CI: 17.3-29.8) and specificity was 100% (95% CI: 99.1-100) with a positive predictive value (PPV) of 100% (95% CI 91.8-100) and a negative predictive value (NPV) of 74.3% (95% CI: 70.5-77.9) as compared to RT-PCR. Conclusions: The high specificity of QuickVue suggest that this POC test can be a useful tool for patient management or triaging during a public health crisis but a low sensitivity suggests that a negative test result need to be further tested using RT-PCR.
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Prevalence of non-responsiveness to an indigenous recombinant hepatitis B vaccine: A study among South Indian health care workers in a tertiary hospital p. 32
RJ Thomas, GJ Fletcher, H Kirupakaran, MP Chacko, S Thenmozhi, CE Eapen, G Chandy, P Abraham
DOI:10.4103/0255-0857.150877  PMID:25657153
Background and Aim: Health care workers (HCW) are at higher risk of contracting HBV infection. Non-response to HBV vaccine is one of the major impediments to prevent healthcare associated HBV infection (HAHI). We estimated the prevalence of non-responsiveness to initial 3-dose regimen of an indigenous recombinant HBV vaccine (GeneVac-B) among South Indian HCWs and typed the HLA in non-responders. Study Design and Method: Of the 778 subjects screened over 1 year, 454 completed all three doses of the hepatitis B vaccination. Anti-HBs titers were estimated by microparticle enzyme immunoassay AxSYM AUSAB, (Abbott, Germany). HLA typing was done using SSP-PCR assay AllSet+™ Gold SSP (Invitrogen, USA). Results: The overall seroconversion rate (anti-HBs > 10 mIU/mL) was 98.89% wherein 90.8% had titers >1000mIU/mL, 7.6% had titers 100-1000mIU/mL, 0.43% had titers < 100 mIU/mL and 1.1% were non-responsive (<10 mIU/mL) to the initial 3-dose regimen. Antibody titers <1000 mIU/mL were significantly associated with the highest quartile of body mass index (BMI) (P < 0.001). We found no significant difference in seroprotection rate between gender (P = 0.088). There was no difference in seroprotection rates among various ethnic groups (P = 0.62). Subjects who were non-responsive in our study had at least one HLA allele earlier known to be associated with non-responsiveness to the vaccine. Conclusion: Our findings suggest that non-response to HBV vaccine is not a major impediment to prevent HAHI. Robust seroprotection rates can be achieved using this indigenous HBV vaccine. However, gender and BMI might influence the level of anti-HBs titers. We recommend the use of this cost effective HBV vaccine as well as postvaccination anti-HBs testing to prevent HAHI among HCWs.
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Optimisation and analysis of polymerase chain reaction based DNA sequencing for genotyping polyoma virus in renal transplant patients: A report from South India p. 37
HN Madhavan, R Bagyalakshmi, M Revathy, P Aarthi, J Malathi
DOI:10.4103/0255-0857.150878  PMID:25657154
Purpose: To optimise a polymerase chain reaction (PCR) based DNA sequencing technique for genotyping polyoma virus in clinical specimens obtained from renal transplant patients. Materials and Methods: A hundred and thirty (106 peripheral blood and 24 urine) clinical specimens collected from renal transplant patients were included in the study for detecting the presence of  DNA of BK virus (BKV), JC virus (JCV) by PCR targeting the viral protein 1 (VP1) gene. PCR based DNA sequencing was performed to determine the genotypes of polyoma virus and subjected to bioinformatics analysis to determine the amino acid sequences and screen for mutations in the VP1 gene. Results: Polyoma virus was detected in 23 (17.69%) specimens of which 19 (82.60%) were positive for BK virus, 3 (13.04%) for JC virus and 1 for both BK and JC virus. PCR based DNA sequencing detected BK virus genotype I in 12 (50%), genotype IV in 8 (33.3%) and JC virus in 4 (16.6%) clinical specimens. BKV genotype I was the predominant genotype (64.2% in peripheral blood and 33.33% in urine) prevalent in south India. Six novel mutations were found - at position 29, 30 to 47 of BKV genotype I; at position 11 and 15 of BKV genotype IV and at position 2 and 30 of JCV. Conclusion: BKV genotype I is the prominent genotype in India and novel mutations detected in the VP1 gene of BKV and JCV are being reported for the first time in literature.
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Association of mycobacterium tuberculosis in the causation of Eales' disease: An institutional experience p. 43
Rajpal , UB Singh, S Mohapatra, VK Wagh, C Porwal, A Kaushik
DOI:10.4103/0255-0857.148829  PMID:25657155
Background: Eales' disease is an idiopathic retinal vasculitis characterized by retinal inflammation, ischemia, and neo-vascularisation. It frequently causes massive vitreous haemorrhage and retinal detachment leading to blindness. Although the exact etiology is unknown, this condition is considered to be a consequence of hypersensitivity reaction to tubercular protein due to previous Mycobacterium tuberculosis (M. tuberculosis) infection. This study is aimed at the detection of association of M. tuberculosis in patients with Eales' disease. Materials and Methods: A prospective case-control study was undertaken in 65 clinically diagnosed cases of Eales' disease. Patients with proliferative diabetic retinopathy, neo-vascular proliferation, macular oedema, premacular fibrosis and tractional retinal detachment were taken as controls. M. tuberculosis DNA was detected (MPT64 gene by polymerase chain reaction, PCR) in patients with Eales' disease. Clinical symptoms along with tuberculin skin test (TST) and erythrocyte sedimentation rate (ESR) were used as gold standard for comparing results of PCR. Result: PCR positivity was found in 12 (38.7%) patients with Eales' disease. The PCR positivity was significantly associated with the patients with high TST reading and high ESR values. Conclusion: Patients with a high TST reading and ESR value and a positive PCR in vitreous samples have a high likelihood of having M. tuberculosis as an etiology.
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Early detection of multi-drug resistance and common mutations in Mycobacterium tuberculosis isolates from Delhi using GenoType MTBDRplus assay p. 46
R Singhal, VP Myneedu, J Arora, N Singh, M Bhalla, A Verma, R Sarin
DOI:10.4103/0255-0857.150879  PMID:25657156
Purpose: There is scarcity of prevalence data of multi-drug-resistant tuberculosis (MDR-TB) data and common mutations responsible in North India. This study aimed to detect MDR-TB among MDR-TB suspects from Delhi and mutation patterns using GenoType MTBDRplus assay. Materials and Methods: All MDR suspects in five districts of New Delhi were referred to the laboratory from 1 st October 2011 to 31 st December 2012 as per criterion defined by Programmatic Management of Drug Resistant Tuberculosis (PMDT). GenoType MTBDRplus assay was performed on 2182 samples or cultures and mutations in the rpoB gene for rifampicin (RIF) and katG and inhA genes for isoniazid (INH) were analyzed. Results: A total of 366 (16.8%) MDR-TB cases were diagnosed. MDR rate was found to be 32%, 16.6% and 10.2% during criterion A, B and C respectively. The most common mutation detected for RIF was S531L (59.0%) and for INH was S315T1 (88.3%). Mutations S531L and S315T1 occurred significantly higher in MDR strains as compared to RIF mono-resistant and INH mono-resistant strains, respectively. Average laboratory turn-around time (TAT) for dispatch of result to districts for test conducted on samples was 4.4 days. Conclusion: GenoType MTBDRplus is a useful assay for rapid detection of MDR-TB. The common mutations for RIF and INH were similar to those seen in other regions. However, mutations determining MDR strains and mono-resistant strains differed significantly for both RIF and INH.
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A method for early detection of antibiotic resistance in positive blood cultures: Experience from an oncology centre in eastern India Highly accessed article p. 53
G Goel, D Das, S Mukherjee, S Bose, K Das, R Mahato, S Bhattacharya
DOI:10.4103/0255-0857.150883  PMID:25657157
Purpose: For antibiotic susceptibility results, conventional culture and sensitivity methods takes 48 hours after a blood culture is flagged positive by automated systems. Early initiation of targeted antibiotic therapy is essential for effective management of sepsis to reduce morbidity, mortality, cost of treatment and prevent antibiotic resistance. Objective of this study was to evaluate Direct Sensitivity Test (DST) as a potential tool to get reliable antibiotic susceptibility results 24 hours earlier. Materials and Methods: Blood cultures flagged positive between May 2011 to December 2012 by BacT/ALERT were Gram stained. All uni-microbial gram-negative blood cultures were simultaneously cultured and processed for DST from broth using disk diffusion method using British Society of Antimicrobial Chemotherapy (BSAC) guidelines. DST results available next day were compared with conventional antibiotic susceptibility test (AST) performed by Vitek-2 on isolated colonies. Results of DST (test method) and AST (reference method) were compared for agreements or errors. Results: Of the 840 antibiotic gram-negative organism combinations tested, Categorical and essential agreements were 83.7% and 96.2% respectively. Minor, major and very major errors were 12.5%, 3.33% and 0.47%, respectively. Conclusions: DST using disk diffusion from positive blood culture broths helps to initiate early targeted antibiotic therapy. There is high concordance between DST and AST.
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COMMENTARY Top

The last mile problem p. 59
B Chatterjee
DOI:10.4103/0255-0857.150886  PMID:25657158
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ORIGINAL ARTICLES Top

The association between serum YKL-40 levels, mean platelet volume, and c-reactive protein in patients with cellulitis p. 61
A Erturk, E Cure, MC Cure, E Parlak, A Kurt, S Ogullar
DOI:10.4103/0255-0857.150891  PMID:25657159
Background: Lower limb cellulitis is an infectious disease that has serious complications unless it is treated. Objectives: In this pilot study, we evaluated whether levels of YKL-40, an acute-phase reactant, and mean platelet volume (MPV), which occurs secondary to inflammation in cellulitis, increase compared to healthy subjects. We also aimed to investigate the association between YKL-40 and MPV in the prognosis of the patients. Material and Methods: A total of 55 patients with cellulitis (23 men and 32 women) and a similar age group of 46 healthy individuals (22 men and 24 women) were included in the study. Cellulitis was diagnosed according to guideline. Serum YKL-40 levels, MPV, C-reactive protein (CRP), and other biochemical values of both groups were compared. Results: YKL-40 levels (52.2 ± 34.5 ng/mL vs 34.6 ± 18.0 ng/mL, P = 0.004), MPV (7.7 ± 1.0 fL vs 6.9 ± 0.7 fL, P < 0.001), and CRP (9.5 ± 8.2 mg/dL vs 0.7 ± 0.6 mg/dL, P < 0.001) were significantly higher in the patients with cellulitis than the control. The mean recovery time (RT) of the patients was 22.6 ± 6.9 days. We found that YKL-40 (odds ratio [OR] 0.1, confidence interval [Cl] 0.028-0.191, P = 0.009) and MPV (OR 2.4, Cl 0.254-4.578, P = 0.029) have an independent association with RT. Conclusion : YKL-40 and MPV values were correlated with higher CRP in the cellulitis group than in controls. According to these results, increased YKL-40 and MPV levels might be a prognostic factor for cellulitis in patients.
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Screening of the novel colicinogenic gram-negative rods against pathogenic Escherichia coli O157:H7 p. 67
H Mushtaq, J Bakht, N Bacha
DOI:10.4103/0255-0857.150895  PMID:25657160
Purpose: Escherichia coli (E. coli) O157:H7 is gram-negative enteric pathogen producing different types of Shiga toxin. This bacterium is the most corporate cause of haemorrhagic colitis in human. Administration of antibiotics (particularly sulfa drugs) against this pathogen is a debatable topic as this may increase the risk of uremic syndrome; especially in children and aged people. Around the world, microbiologists are in search of alternative therapeutic methods specially probiotics against this pathogen. In the present study, we have focused on the investigation of alternate bio-therapeutics (probiotics) for the treatment of patients infected with E. coli O157:H7. This study is based on the identification of colicin-producing gram-negative bacteria (particularly enterobacteriaceae) which can competently exclude E. coli O157:H7 from the gut of the infected individual. Materials and Methods: Hundred samples from human, animal faeces and septic tank water were analysed for nonpathogenic gram-negative rods (GNRs). Results: Out of these samples, 175 isolates of GNRs were checked for their activity against E. coli O157:H7. Only 47 isolates inhibited the growth of E. coli O157:H7, among which majority were identified as E. coli. These E. coli strains were found to be the efficient producers of colicin. Some of the closely related species i. e., Citrobacter sp, Pantoea sp. and Kluyvera sp. also showed considerable colicinogenic activity. Moreover, colicinogenic species were found to be nonhaemolytic, tolerant to acidic environment (pH 3) and sensitive to commonly used antibiotics. Conclusion: Nonhaemolytic, acid tolerant and sensitive to antibiotics suggests the possible use of these circulating endothelial cells (CEC) as inexpensive and inoffensive therapeutic agent (probiotics) in E. coli O157:H7 infections.
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Molecular characterization of CTX-M type Extended Spectrum Beta Lactamase producing E. coli isolated from humans and the environment p. 73
EA George, S Sankar, MV Jesudasan, C Sudandiradoss, B Nandagopal
DOI:10.4103/0255-0857.150896  PMID:25657161
Purpose: Escherichia coli is a common pathogen causing community- and hospital-acquired infections. The infections caused by the Extended Spectrum Beta Lactamase (ESBL) enzymes-producing E. coli hinder antibiotic treatment. Materials and Methods: Plasmid DNA samples were subjected to PCR specific for TEM, SHV and CTX-M genes obtained from 110 E. coli strains isolated from hospitalized patients, healthy individuals and environment in Vellore, South India . Results: Among the 110 isolates tested, 21.8% were positive for TEM and 2.7% positive for SHV and 91.8% positive for CTX-M. The proportion of CTX-M positive E. coli was not statistically different between the study groups. Nineteen of 20 strains were CTX-M-15 type and the other was CTX-M-14 type. The phylogenetic analysis of 19 strains clustered with the pandemic CTX-M-15-ST131 strain, indicating this as an evolving global problem for antibiotic therapy. The geomapping of samples indicated 'hotspot' areas of healthy individuals, patients and the environmental samples. Conclusion: The spatial presentation of GIS mapping allowed identification of clustering among patients and healthy individuals and contaminated environmental points.
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Molecular characterization of Enterococcus spp. clinical isolates from Cairo, Egypt p. 80
YA Hashem, AS Yassin, MA Amin
DOI:10.4103/0255-0857.148836  PMID:25657162
Purpose: Enterococci are responsible for serious diseases such as bacteraemia, endocarditis and urinary tract infections. The ability of enterococci to cause such diseases is due to acquisition of certain virulence factors such as haemolysin, gelatinase and enterococcus surface protein. This study has been conducted to investigate the occurrence of virulence factors and resistance to various antibiotics with emphasis on vancomycin in the Enterococcus spp. Materials and Methods: Clinical specimens were collected and isolates were identified by proper microscopic, culture and biochemical tests. Susceptibility and degree of resistance of the isolates to various antibiotics were determined. Virulence factors were examined by phenotypic tests followed by molecular methods. Bioinformatics analysis was used to detect regions in the genomes that might have originated from horizontal gene transfer. Result: The presence or absence of virulence genes did not affect the pattern of antimicrobial resistance in Enterococcus isolates; consequently, no relationship was found between virulence factors and resistance to different antibiotics used. Bioinformatics analysis showed that the virulence genes were mainly transferred by transposons. Conclusion: Among the enterococci, environmental factors may interfere in the expression of virulence factors. Horizontal gene transfer plays an important role in the spread of resistance and virulence genes.
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The dissemination mode of drug-resistant genes in Enterobacter cloacae p. 87
J Liu, T Zeng, G Su, LY Lin, Y Zhao, WQ Yang, WX Xie, ZG Zhao, GM Li
DOI:10.4103/0255-0857.150899  PMID:25657163
Background: Enterobacter cloacae (E. cloacae) infection has the highest mortality rate among Enterobacter infections. This study aimed to determine the prevalence and the transmission route of the class I integron, qnr genes, and CTX-M ESBLs genes in clinical isolates and to analyse the association between the prevalence of MDR genes and the antibiotic resistance of E. cloacae. Materials and Methods: The antibiotic susceptibility was tested the agar dilution method. The class I integron, qnr genes, and CTX-M ESBLs genes were detected by polymerase chain reaction (PCR). The prevalence data were analysed with the Chi-square test. Results: In the 100 clinical isolates, the class I integron-positive rate was 65%, with 12% on chromosome, 15% on plasmids and 38% on both. The positive rate of qnr genes was 37% with plasmid location. The positive rates for qnrA, qnrB and qnrS were 6%, 23% and 8%, respectively. The CTX-M ESBLs-positive rate was 34%. For CTX-M-1 ESBLs, 15% were on chromosome, 6% on plasmids and 4% on both; for CTX-M-9 ESBLs, 1% was on chromosome and 7% on plasmid; for CTX-M-25 ESBLs, 3% were on chromosome and 1% on plasmid. Conclusion: Antibiotic resistance genes may be horizontally and vertically disseminated among E. cloacae, which helps multidrug-resistant (MDR) strains of E. cloacae to be successful nosocomial agents.
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Susceptibility characterisation of Candida spp. to four essential oils p. 93
CC Rath, S Mohapatra
DOI:10.4103/0255-0857.150903  PMID:25657164
In the present investigation, anti-Candida activity of four essential oils i.e. Black cumin (Nigella sativa), Curry leaf (Murraya koienigii), Ajwain (Trachiyspirum ammi), and Betel leaf (Piper betel) were screened against four human pathogenic species of Candida viz. Candida albicans, Candida tropicalis, Candida glabrata, and Candida parapsilosis. The minimum inhibitory concentration (MIC) values of the oils ranged between 15.62 and 250 μl/ml while studied through tube dilution method. The oils retained their anti-Candida activities even after heat treatment (at 45ΊC, 60ΊC, 100ΊC for 1 hour) and also on autoclaving. Both Ajwain and Black Cumin leaf oils showed better anti-Candida activity against Candida albicans, resulting in an irreversible damage to the cells. The anti-Candida activity of these essential oils could be attributable to the membrane inhibition mechanism. The activity of the oils is reported to be microbicidal (Candida-cidal).
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Effects of fluconazole treatment of mice infected with fluconazole-susceptible and -resistant Candida tropicalis on fungal cell surface hydrophobicity, adhesion and biofilm formation p. 97
RL Kanoshiki, SB de Paula, JP Santos, AT Morey, NB Souza, LM Yamauchi, BP Dias Filho, SF Yamada-Ogatta
DOI:10.4103/0255-0857.148834  PMID:25657165
Background : The incidence of Candida tropicalis less susceptible to fluconazole (FLC) has been reported in many parts of the world. Objectives : The aim of this study was to examine the changes of putative virulence attributes of Candida tropicalis accompanying the development of resistance to FLC in vitro and in vivo. Materials and Methods : A FLC-resistant strain (FLC-R) was obtained after sequential exposure of a clinical isolate FLC-sensitive (FLC-S) to increasing concentrations of the antifungal. The course of infection by both strains was analyzed in BALB/c mice. Analyses of gene expression were performed by real-time polymerase chain reaction PCR. The cell surface hydrophobicity, adhesion and biofilm formation were also determined. Results : Development of resistance to FLC could be observed after 15 days of subculture in azole-containing medium. Overexpression of MDR1 and ERG11 genes were observed in FLC-R, and this strain exhibited enhanced virulence in mice, as assessed by the mortality rate. All mice challenged with the FLC-R died and FLC-treatment caused earlier death in mice infected with this strain. All animals challenged with FLC-S survived the experiment, regardless of FLC-treatment. Overall, FLC-R derivatives strains were significantly more hydrophobic than FLC-S strains and showed greater adherence and higher capacity to form biofilm on polystyrene surface. Conclusions : The expression of virulence factors was higher in FLC-R-C. tropicalis and it was enhanced after FLC-exposure. These data alert us to the importance of identifying microorganisms that show resistance to the antifungals to establish an appropriate management of candidiasis therapy.
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Prevalence of Hepatitis A virus (HAV) and Hepatitis E virus (HEV) in the patients presenting with acute viral hepatitis p. 102
A Joon, P Rao, SM Shenoy, S Baliga
DOI:10.4103/0255-0857.150908  PMID:25657123
Background: Hepatitis A virus (HAV) and Hepatitis E virus (HEV) are both enterically transmitted, resulting in acute viral hepatitis (AVH) in developing countries. They pose major health problems in our country. This study was done to determine prevalence of HAV and HEV in patients presenting with AVH and the co-infection of HAV and HEV in these patients. Materials and Methods: A cross-sectional study of 2-years duration was conducted in the Department of Microbiology, KMC, Mangalore. A non-random sampling of 958 patients presenting with AVH was considered in the study. On the basis of history, serum samples were analysed for IgM anti-HAV and IgM anti-HEV for the detection of HAV and HEV, respectively using commercially available ELISA kits. Data collected was analysed by using Statistical Package for the Social Sciences (SPSS) version 11.5. Results: The seroprevalence of HAV- and HEV-positive patients were 19.31% and 10.54%, respectively. The seroprevalence of both HAV and HEV in patients with acute viral hepatitis was 11.5%. The prevalence of HAV and HEV among males (68% and 31%) was higher than in females (31% and 20%) and was predominantly seen among young adults. These infections were predominantly seen during end of monsoons and beginning of winter. Conclusion: Though the prevalence of HAV is much higher than that of HEV, co-infection rate of 11.5% mandates the screening for HEV which will be of immense importance in pregnant women and improving levels of personal hygiene among higher socio-economic population. These data will be essential for planning of future vaccination strategies and for better sanitation programme in this part of the country.
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Utility of routine real time quantitative PCR monitoring of HCV infection in haemodialysis patients p. 106
S Datta, N Goel, C Wattal
DOI:10.4103/0255-0857.148832  PMID:25657124
Background: Hepatitis C virus (HCV) infection causes significant morbidity and mortality in patients of end stage renal disease (ESRD) on haemodialysis (HD). Stringent screening methods can help in its early diagnosis. Objective: The study addresses the utility of real-time quantitative polymerase chain reaction (RQ-PCR) in the diagnosis and monitoring of HCV infection especially on seronegative and normal serum alanine aminotransferase (ALT) HD patients. Material and Methods: This retrospective study was carried out from January 2010 to December 2012. Patients of ESRD on maintenance HD and on whom all the three assays HCV antibody serology, PCR and ALT were done were included in the study (n = 123). Group 1 (n = 57), comprised of patients with negative serology and normal ALT, and Group 2 (n = 66), had either raised ALT and or a positive or equivocal serology. Results: Out of the 123 cases studied, HCV serology was positive in 36.5% (45), ALT raised in 18.6% (23) and PCR positive in 67.4% (83) cases. PCR positivity was significantly higher than serology and raised ALT. Group 2 had a significantly higher PCR positivity than Group 1 (P = 0.0004), but 50.9% patients of Group 1, were also PCR positive and 69% of them had a high viral count of >8 × 10 5 IU/ml at the time of detection. Conclusion: Regular routine screening of HCV by RQ-PCR in ESRD patients can help in early diagnosis of HCV infection in patients with low index of suspicion.
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Biofilm as a virulence marker in Candida species in Nosocomial blood stream infection and its correlation with antifungal resistance p. 112
M Bhatt, G Sarangi, BP Paty, D Mohapatra, N Chayani, A Mahapatra, P Das, D Sahoo
DOI:10.4103/0255-0857.150909  PMID:25657126
Nosocomial blood stream infections (BSI) due to fungi especially Candida is increasing steadily. A two year prospective study was conducted in the S.C.B. Medical College with an aim to evaluate the species distribution, antifungal susceptibility and biofilm formation of Candida spp. isolated from nosocomial BSIs. 34 Candida spp. were isolated from 359 blood cultures. Antifungal susceptibility was performed by microbroth dilution technique and both visual and spectrophotometric method were used for biofilm detection. C. tropicalis was the common spp. isolated followed by C. parapsilosis and others. Most (92%) of the isolates were susceptible to Amphoterecin-B and highest resistance was observed against Flucytosine (37%) and Fluconazole(35%). Biofilm production and antifungal resistance was observed more in nonalbicans Candida spp.
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Microbial colonisation of orthopaedic tourniquets: A potential risk for surgical site infection p. 115
SK Sahu, B Tudu, PK Mall
DOI:10.4103/0255-0857.150910  PMID:25657127
Pneumatic tourniquets have been used in orthopaedic surgery to get avascular fields. Sixteen such tourniquets were analysed for microbial colonisation. Samples were taken from two inner and two outer areas of each tourniquet and cultured on sheep blood agar. Eight of these were wiped with Savlon and the rest with Sterillium solution. Post-treatment samples from the same sites were again cultured. After incubation, colonies from each site were identified and counted. It was observed that the tourniquets were colonised with coagulase-negative staphylococci, Staphylococcus aureus, Bacillus, diphtheroids, Pseudomonas, Acinetobacter, enterococci, enterobacteria, and Candida. On treating with Savlon and Sterillium, there was 92.18% and 95.70% reduction in the colony count, respectively.
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On the path of continual improvement: An evaluation of biomedical waste management training p. 119
K Usha Krishnan, T Sheila Doris Devamani, G Jayalakshmi
DOI:10.4103/0255-0857.150911  PMID:25657128
Microbiologists play a pivotal role in the prevention of infection both at hospital and community level. Biomedical waste management (BMWM) is one of the aspects of infection control. For the effective implementation of this, various training programmes have been conducted at our regional training centre. This study evaluates the impact of the BMWM training programme. Knowledge on all parameters was significantly better at post-course evaluation for all the participants (P value 0.001). Evaluation of training is recommended to improve the effectiveness of training method.
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Extra pulmonary tuberculosis: Rapid identification of Mycobacterium tuberculosis grown in Mycobacterium growth indicator tube 960 and Lowenstein-Jensen media, employing Standard diagnostics Bioline Mycobacterium tuberculosis protein 64 antigen detection kit p. 122
G Kandhakumari, S Stephen
DOI:10.4103/0255-0857.150912  PMID:25657129
Background: Investigation of extra pulmonary tuberculosis (EPTB) in and around Pondicherry is being carried out since August 2011 in our tertiary care super specialty hospital. Objectives: To compare the rapid Kit SD Bio-Line MPT 64 Ag with conventional and time consuming biochemical tests. Confirmation of Mycobacterium tuberculosis at a reasonable time frame is the main thrust. Materials and Methods: Thirty three Mycobacterium tuberculosis and four Non-Tuberculous Mycobacteria (NTM) grown in MGIT960 system/Lowenstein-Jensen media (LJ) were examined by the rapid MPT 64 antigen detection as well as a battery of conventional tests like niacin, nitrate reduction, paraminobenzoic acid susceptibility and cord formation. Results and Conclusion: . Both the rapid kit and conventional tests correctly identified 33 M.tuberculosis isolates. Keeping conventional identification as reference, sensitivity and specificity for rapid kit was 100%. Rapid kit which takes only 15 minutes is accurate, cost effective, and facilitates early treatment for these EPTB patients, whose clinical specimens are paucibacillary.
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Declining trends of Syphilis seroprevalance among antenatal clinic cases and STD clinic cases in a tertiary care centre: From January 2002 to December 2012 p. 126
S Chopra, A Garg, M Chopra, A Ghosh, V Sreenivas, S Sood, A Kapil, BK Das
DOI:10.4103/0255-0857.150917  PMID:25657130
Syphilis affects nearly 1.36 million pregnant women, majority of these cases being concentrated in the developing countries. We aimed at analysing the 11 years' trends in syphilis seroprevalence in antenatal clinics (ANC) and STD clinic cases. Laboratory data of syphilis cases from 2002-2012 were retrospectively analysed. Out of the total 73642 cases, 393 (0.53%) tested positive for Syphilis. A statistically significant decline in syphilis prevalence was found in both ANC and STD clinic cases. The efforts of various interventional programs should continue to make the screening and treatment facilities readily accessible to continue the decline in syphilis seropositivity.
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ORIGINAL ARTICLE Top

Molecular epidemiology of methicillin resistant staphylococcus aureus colonizing the anterior Nares of school children of Udupi Taluk p. 129
S Govindan, AS Maroli, AM Ciraj, I Bairy
DOI:10.4103/0255-0857.150919  PMID:25657131
Context: Community associated methicillin resistant Staphylococcus aureus (CA-MRSA) cause serious skin and soft tissue infections including necrotizing fasciitis and necrotizing pneumonia. Production of Panton Valentine Leucocidine (PVL) toxin is implicated in its enhanced virulence. A variant of epidemic MRSA-15 (EMRSA-15) which produces PVL toxin has been isolated and characterized by pulsed-field gel electrophoresis (PFGE) method from the Indian population both in hospital and community settings. Aims: Identify the epidemiological type of MRSA colonizing the anterior nares of school children in Udupi taluk. Settings and Design: The study population included children of the age group of 5-16 years belonging to the Udupi taluk of Karnataka, India. A total of 1503 children were screened for MRSA colonization during July 2009 to December 2010. Materials and Methods: PVL assay, Staphylococcal Cassette Chromosome (SCC) mec typing and PFGE typing were carried out with all the MRSA isolates. Statistical Analysis Used: Frequency distribution of different variables was assessed by SPSS. Results: Among the 1.1% of MRSA, 58.8% (10/17) of isolates were positive for pvl and 41.7% (7/17) were identified as SCC mec type IV. PFGE patterns of all the strains were identical with Indian variant EMRSA-15; however they were different from classical EMRSA-15 in 3-4 bands. Conclusions: The Indian variant EMRSA-15 gains much epidemiological relevance owing to the acquisition of pvl gene. In spite of low prevalence of nasal colonization of MRSA, emergence of the virulent Indian variant EMRSA-15 in our community is a worrisome fact to be reckoned with.
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Epidemiological and clinical pattern of dermatomycoses in rural India p. 134
A Lakshmanan, P Ganeshkumar, S Raam Mohan, M Hemamalini, R Madhavan
DOI:10.4103/0255-0857.150922  PMID:25657132
Superficial fungal infections are most common in tropical and subtropical countries. In this study, 297 suspected superficial fungal infection cases were identified among 15,950 patients screened. The collected samples (skin, nail, and hair) were subjected to direct microscopy with potassium hydroxide and cultured on Sabourauds dextrose agar to identify the fungal species. The prevalence of superficial fungal infection was 27.6% (82/297), dermatophytosis was 75.6% (62/82), and non-dermatophytosis was 24.4% (20/82). Among the isolated dermatophytes, Trichophyton rubrum was the commonest species (79%) and Candida (60%) the commonest non-dermatophytic species. Tinea corporis was the commonest (78%) clinical presentation.
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CASE REPORTS Top

Suppurative supraclavicular bacille calmette-guerine lymphadenitis - A case report, awareness and management options p. 137
US Udgaonkar, SS Patil, VB Rekha, S Shah
DOI:10.4103/0255-0857.150928  PMID:25657133
Diagnosis of Bacille calmette-guerine (BCG) adenitis is clinical. Conventional laboratory tests do not differentiate BCG adenitis from tuberculous adenitis. We report a case of a 3-month-old healthy baby presenting with suppurative BCG adenitis. FNAC revealed AFB on ZN-Staining, later confirmed to be Mycobacterium bovis by multiplex PCR. The treatment of suppurative BCG adenitis is needle aspiration. Anti-tubercular treatment is unwarranted.
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Mother to child transmission of hepatitis B virus: A cause for concern p. 140
AM Ismail, A Raghavendran, J Sivakumar, M Radhakrishnan, W Rose, P Abraham
DOI:10.4103/0255-0857.150931  PMID:25657134
Mother to child transmission of hepatitis B virus (HBV) is an important public health issue. India introduced HBV vaccine in 10 states as part of its Universal Immunization Program (UIP). Here we show evidence of mother-to-child transmission of HBV in three families from Jharkhand and Bihar states where HBV vaccination is not yet included in the UIP. This report illustrates the need for active screening of HBV in pregnant women and implementation of HBV vaccine across all states in India to reduce the burden of disease.
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Spontaneous clearance of chronic hepatitis C infection is associated with an internal ribosomal entry site IIId stem loop structure variant p. 143
NG Bader El Din, MM El Hefnawy, MH Omran, RM Dawood, Y El Abd, MK Ibrahim, MK El Awady
DOI:10.4103/0255-0857.148835  PMID:25657135
Aim: To investigate if any mutations in hepatitis C virus (HCV) internal ribosome entry site (IRES) can inhibit the translation of viral polyprotein. Materials and Methods: A 26-year-old male patient infected with HCV 10 years ago was followed up. After 9 years of chronic infection. The patient had managed to resolve the infection for a period of 9 months, after which the patient experienced a viral recurrence characterized by high viral load and diverse HCV quasispecies. The IRES structures of the viral strains that disappeared were comparable with those that are currently active using structural mutational analysis. Results: A novo mutational position 254 combined with a rarely observed mutation at position 253 in the stem of the IIId subdomain were observed and the new conformation had an octa-apical loop (AGUGUUGG) and a shift in the 3 ` GU from the loop to the stem. Conclusions: These mutations were found to be highly deleterious, and they affected the direct binding of the IIId loop to the 40S ribosomal subunit with a subsequent inhibition of translation of viral polyprotein and clearance of the virus.
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Fatal cryptococcosis involving multiple sites in an immunocompetent child p. 148
H Kaur, K Zaman, BR Thapa, SM Rudramurthy
DOI:10.4103/0255-0857.150935  PMID:25657136
Disseminated cryptococcosis is less common in immunocompetent individuals. Herein, we report a fatal case of cryptococcosis in apparently immunocompetent child with multiple site involvement. The yeast isolated from cerebrospinal fluid, blood, endotracheal, gastric and lymph node aspirate was identified by molecular method as Cryptococcus neoformans var. grubii.
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Multiplex PCR in diagnosis of M. tuberculosis and M. avium co-infection from lymph node in an AIDS patient p. 151
K Sharma, A Mewara, N Gupta, A Sharma, S Varma
DOI:10.4103/0255-0857.150940  PMID:25657138
A 35-year-old, HIV-seropositive male (CD4 count 41 cells/mm 3 ) on highly active antiretroviral ( HAART) presented with fever and weight loss for 3 months and new skin lesions. He was earlier diagnosed of TB and was on anti-tubercular therapy (ATT). The retroperitoneal lymph node aspirate showed acid-fast bacilli and epithelioid cell granulomas; however, cultures remained sterile. A dual infection with Mycobacterium tuberculosis and Mycobacterium avium was diagnosed with multiplex polymerase chain reaction (MPCR). Clarithromycin was added to ATT, and on follow-up at 1 and 3 months, the patient responded well. Molecular methods like MPCR should be exploited for routine diagnosis of high-risk patients.
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Chikungunya virus infection amongst the acute encephalitis syndrome cases in West Bengal, India p. 153
D Taraphdar, BK Roy, S Chatterjee
DOI:10.4103/0255-0857.150946  PMID:25657139
Chikungunya virus (CHIKV) infection from the acute encephalitis syndrome cases is an uncommon form and has been observed in the year 2010-11 from West Bengal, India. The case-1 and case-2 had the acute encephalitis syndrome; case-3 was of acute disseminated encephalomyelitis whereas the case-4 had the symptoms of meningo-encephalopathy with bulbar involvement. We are reporting four cases with neurological complications involving central nervous system (CNS) due to CHIKV infection from this state for the first time. The virus has spread almost every districts of this state rapidly. At this stage, these cases are public health threat.
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First case of Paragonimus westermani infection in a female patient in India p. 156
TS Singh, S Hiromu, KR Devi, WA Singh
DOI:10.4103/0255-0857.150950  PMID:25657140
Paragonimiasis is a foodborne parasitic zoonosis caused by lung fluke species of the genus Paragonimus. The Paragonimus westermani is the most common human pathogen in Asian countries. In northeast India, Paragonimus heterotremus has been documented as the only human pathogen in the earlier literature. In India, P. westermani infection in humans remained undetermined. Herein, we report a case of pulmonary paragonimiasis due to P. westermani in an adult female in Manipur. The diagnosis was made by morphological and molecular characterisation of the eggs in the sputum. This is the first confirmed case of paragonimiasis due to P. westermani in India.
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CORRESPONDENCE Top

Report of carbapenem resistant Ps. aeruginosa, isolates carrying ESBLs, AmpC and MBL enzymes based on phenotypic methodology and susceptibility to Fosfomycin p. 160
R Garg, V Gupta, J Chander, M Kaur
DOI:10.4103/0255-0857.150954  PMID:25657141
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Detection and distribution pattern of prevalent genotypes of Hepatitis-C virus among chronic hepatitis patients from Kumaon region of Uttarakhand, India p. 161
YPS Malik, N Kumar, V Rawat, K Sharma, A Kumar, M Singhai, M Kumar
DOI:10.4103/0255-0857.148838  PMID:25657142
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Mucor -culture/molecular analysis necessary for identification p. 163
S Duggal, SR Rongpharpi
DOI:10.4103/0255-0857.150960  PMID:25657143
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In vitro activities of linezolid against clinical isolates of Mycobacterium tuberculosis from Shenyang, north of China p. 164
H Liu, Y Wang, N Liu, L Zhao
DOI:10.4103/0255-0857.150967  PMID:25657144
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Primary thoracic wall tuberculosis: A rare case p. 165
P Sharma, VK Mehta, RC Guleria, D Singh, A Kanga, S Mohindroo
DOI:10.4103/0255-0857.150971  PMID:25657145
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Vancomycin-resistant enterococcus (VRE) vs Methicillin-resistant Staphylococcus Aureus (MRSA) p. 166
KS Ashwin, NP Muralidharan
DOI:10.4103/0255-0857.150976  PMID:25657146
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Need of microbiological investigations in the case of infected hydatid cyst p. 168
US Udgaonkar, SR Shah
DOI:10.4103/0255-0857.150981  PMID:25657147
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Bacillus cereus bacteraemia in a patient of acute myeloid leukaemia p. 168
Ratnamani Sharma, Ratna Rao
DOI:10.4103/0255-0857.150982  PMID:25657148
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SNIPPETS Top

Research Snippets p. 171
P Desikan
DOI:10.4103/0255-0857.150984  PMID:25657149
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2004 - Indian Journal of Medical Microbiology
Published by Wolters Kluwer - Medknow

Online since April 2001, new site since 1st August '04