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   2010| July-September  | Volume 28 | Issue 3  
    Online since July 17, 2010

 
 
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CORRESPONDENCE
New Delhi metallo-beta-lactamases: A wake-up call for microbiologists
BVS Krishna
July-September 2010, 28(3):265-266
DOI:10.4103/0255-0857.66477  PMID:20644324
  17 15,649 1,496
ORIGINAL ARTICLES
Prevalence and risk factors for colonisation with extended spectrum β-lactamase producing enterobacteriacae vis-à-vis usage of antimicrobials
JB Sarma, GU Ahmed
July-September 2010, 28(3):217-220
DOI:10.4103/0255-0857.66476  PMID:20644309
Purpose: A point prevalence study was carried out in a teaching hospital in Assam to determine the prevalence, sensitivity profile and risk factors for acquisition of extended spectrum β-lactamase (ESBL) producing enterobacteriacae vis-ΰ-vis amount and pattern of antibiotic use. Materials and Methods: ESBL was detected by double disc synergy method. Defined daily dose and bed-days were calculated. Result: Colonisation rate of ESBL producing enterobacteriacae ranged from 14% (n=73) in medicine to the highest 41% (n=29) in orthopaedic with an intermediate 23% (n=80) in surgery. Presence of ESBL was found to be strongly associated with resistance to specific classes of antimicrobials. Exposure to cefotaxime and gentamicin, and surgery were risk factors for acquiring ESBL producing enterobacteriacae. Non-ESBL producing community isolates were found to be considerably more sensitive to different antibiotics with no resistance detected to trimethoprim, co-trimoxazole, ciprofloxacin and aminoglycosides. Conclusion: The study confirms the role of certain 'high risk' antimicrobials in acquisition of ESBL producing Enterobacteriaceae and shows that periodic cohort studies could be an effective strategy in surveillance of antimicrobial resistance in hospitals of resource poor countries to inform antibiotic policy and treatment guidelines.
  7 4,369 617
EDITORIAL
National antibiotic resistance surveillance and control
D Raghunath
July-September 2010, 28(3):189-190
DOI:10.4103/0255-0857.66463  PMID:20644302
  6 6,189 1,097
ORIGINAL ARTICLES
Evaluation of a nested PCR targeting IS6110 of Mycobacterium tuberculosis for detection of the organism in the leukocyte fraction of blood samples
B Nandagopal, S Sankar, K Lingesan, KC Appu, G Sridharan, AK Gopinathan
July-September 2010, 28(3):227-232
DOI:10.4103/0255-0857.66480  PMID:20644311
Purpose: Tuberculosis poses a serious health problem in resource-poor settings such as India. Polymerase chain reaction (PCR) is presently seen as a promising alternative to conventional smear microscopy and culture techniques. Undiagnosed fever is a condition where the aetiology could include tuberculosis in a significant percentage. This paper evaluates a nested PCR (nPCR) using Hotstar Taq for the detection of M. tuberculosis in patients with febrile illness using insertion element, IS6110 as a target. Material and Methods: A total of 355 samples (301 HIV status unknown and 54 HIV seropositives) from patients primarily with febrile illness were tested for the presence of M. tuberculosis. Blood culture was done in a commercial automated blood culture system and nPCR in DNA extracts from buffy coat samples. Hotstar Taq polymerase was used to enhance the sensitivity of nPCR and the lower limit of detection was determined by using cloned plasmid. Results: Among the patients tested, 2% were positive by automated culture system and 6.8% of patients were positive by nPCR. Majority of the positives were from HIV seropositive individuals. The sensitivity of the nPCR was 100% and the specificity was 95.1%. The lower limit of detection was less than 1 genome copy per microlitre. Among the nPCR positives, patients from rural community were significantly higher than from the peri-urban community. Conclusions: The nPCR had a high sensitivity and specificity on buffy coat samples using Hotstar Taq polymerase in the reaction mix. Thus the technique is a valuable tool in the diagnosis of tuberculosis.
  6 6,390 423
Detection of Mycobacterium tuberculosis resistance mutations to rifampin and isoniazid by real-time PCR
A Hristea, D Otelea, S Paraschiv, A Macri, C Baicus, O Moldovan, M Tinischi, V Arama, A Streinu-Cercel
July-September 2010, 28(3):211-216
DOI:10.4103/0255-0857.66474  PMID:20644308
Objective: The objective of our study was to evaluate the use of a real-time polymerase chain reaction (PCR)-based technique for the prediction of phenotypic resistance of Mycobacterium tuberculosis. Materials and Methods: We tested 67 M tuberculosis strains (26 drug resistant and 41 drug susceptible) using a method recommended for the LightCycler platform. The susceptibility testing was performed by the absolute concentration method. For rifampin resistance, two regions of the rpoB gene were targeted, while for identification of isoniazid resistance, we searched for mutations in katG and inhA genes. Results: The sensitivity and specificity of this method for rapid detection of mutations for isoniazid resistance were 96% (95% CI: 88% to 100%) and 95% (95% CI: 89% to 100%), respectively. For detection of rifampin resistance, the sensitivity and specificity were 92% (95% CI: 81% to 100%) and 74% (95% CI: 61% to 87%), respectively. The main isoniazid resistance mechanism identified in our isolates is related to changes in the katG gene that encodes catalase. We found that for rifampin resistance the concordance between the predicted and observed phenotype was less than satisfactory. Conclusions: Using this method, the best accuracy for genotyping compared with phenotypic resistance testing was obtained for detecting isoniazid resistance mutations. Although real-time PCR assay may be a valuable diagnostic tool, it is not yet completely satisfactory for detection of drug resistance mutations in M tuberculosis.
  6 7,563 615
BRIEF COMMUNICATIONS
Detection and characterization of metallo beta lactamases producing Pseudomonas aeruginosa
A Manoharan, S Chatterjee, D Mathai, SARI Study Group
July-September 2010, 28(3):241-244
DOI:10.4103/0255-0857.66486  PMID:20644314
This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs) among nosocomial Pseudomonas aeruginosa. Sixty one among 176 P. aeruginosa isolates, collected as part of a multicentric study (2005-2007), were evaluated for carbapenem resistance (CARB-R; resistant to either imipenem/meropenem) and screened for MBL by Combination Disk Diffusion Test (CDDT) using imipenem (IMP), meropenem (MER) and ceftazidime (CAZ) with EDTA. MBL positives were further confirmed by IMP + EDTA Etest. Twenty strains (42.6%) were found to be MBL producers among the 61 P. aeruginosa. PCR for IMP and VIM MBL was performed on 48 of the 61, 15 were positive for VIM MBL type. CDDT using IMP + EDTA had the highest sensitivity and specificity of 87.8% and 84.4% when compared to Etest, which was higher than the values obtained for CAZ + EDTA and MER + EDTA. CDDT using IMP + EDTA also compared very well with the PCR (specificity = 90.9%, sensitivity = 93.3%). CARB-R among P. aeruginosa is mediated predominantly via MBL production. Clinical P. aeruginosa isolates can be screened routinely using the less expensive IMP + EDTA CDDT in clinical microbiology laboratories.
  5 12,140 1,328
CASE REPORTS
Ophthalmomyiasis: Three cases from North India
S Khurana, M Biswal, HS Bhatti, SS Pandav, A Gupta, SS Chatterjee, WV Lyngdoh, N Malla
July-September 2010, 28(3):257-261
DOI:10.4103/0255-0857.66490  PMID:20644320
Three cases of external ophthalmomyiasis are reported here. The larvae were identified to be Oestrus ovis in two cases and Cochliomyia hominivorax in one. Two of the patients were immunocompetent while one was undergoing treatment for squamous cell carcinoma of eyelid. In the latter myiasis led to complete destruction of the eye.
  5 4,554 205
SPECIAL ARTICLES
Biomedical waste management: Incineration vs. environmental safety
V Gautam, R Thapar, M Sharma
July-September 2010, 28(3):191-192
DOI:10.4103/0255-0857.66465  PMID:20644303
Public concerns about incinerator emissions, as well as the creation of federal regulations for medical waste incinerators, are causing many health care facilities to rethink their choices in medical waste treatment. As stated by Health Care Without Harm, non-incineration treatment technologies are a growing and developing field. Most medical waste is incinerated, a practice that is short-lived because of environmental considerations. The burning of solid and regulated medical waste generated by health care creates many problems. Medical waste incinerators emit toxic air pollutants and toxic ash residues that are the major source of dioxins in the environment. International Agency for Research on Cancer, an arm of WHO, acknowledged dioxins cancer causing potential and classified it as human carcinogen. Development of waste management policies, careful waste segregation and training programs, as well as attention to materials purchased, are essential in minimizing the environmental and health impacts of any technology.
  5 20,831 1,083
ORIGINAL ARTICLES
Observation on integron carriage among clinical isolates of Klebsiella pneumoniae producing extended-spectrum β-lactamases
A Bhattacharjee, MR Sen, P Prakash, A Gaur, S Anupurba, G Nath
July-September 2010, 28(3):207-210
DOI:10.4103/0255-0857.66472  PMID:20644307
Purpose: Klebsiella pneumoniae is considered an important pathogen causing nosocomial and community-acquired infections and is often associated with the production of extended-spectrum β-lactamases (ESBL) belonging to SHV and CTX-M families, which are frequently described as a part of complex integrons, facilitate their horizontal transfer to other related as well as unrelated microbes. The present study was undertaken to investigate the occurrence and characterization of integrons among K pneumoniae isolates producing ESBL in a tertiary referral hospital. Materials and Methods: A total of 136 clinical isolates of K pneumoniae were investigated for the presence of ESBL. Their ESBL genes were characterized by multiplex polymerase chain reaction (PCR). Integrase gene PCR was performed to detect the presence of integron. The isolates were further typed by random amplification of polymorphic DNA (RAPD). Result: Out of 136 K pneumoniae isolates, 63 (46%) were confirmed to be ESBL producers. SHV (68%) and CTX-M (67%) ESBL genes were the most common in our study. Of the 63 ESBL-positive isolates, 58 (92%) strains carried integrons; 52 strains (82%) carried only class 1 integron, whereas 6 (9%) isolates harboured both class 2 integrons and the class 1 gene. However, in ESBL negatives, only 29 (40%) strains were positive for class 1 integron and none for class 2 integron. Conclusion: The presence of class 2 integron amongst ESBL-producing K pneumoniae is being described for the first time in this part of the world. The findings of this study strongly suggest that integrons have a role in the dissemination of ESBL-mediated resistance among the nosocomial isolates of K pneumonia.
  4 4,920 420
CASE REPORTS
Mycobacterium chelonae and Mycobacterium fortuitum infection following open fracture: A case report and review of the literature
K Kwan, ST Ho
July-September 2010, 28(3):248-250
DOI:10.4103/0255-0857.66488  PMID:20644316
We report a case of dual nontuberculous mycobacterial infections complicating an open distal radius and ulna fracture after polytrauma in a 35-year-old man. There was persistent wound discharge after definitive fixation of this fracture, but microbiological cultures did not yield any organism. The patient underwent multiple debridement, and subsequent tissue grew Mycobacterium chelonae and Mycobacterium fortuitum. Despite appropriate chemotherapy and surgical debridement the infection persisted until radical bone excision and tissue debridement were done. This case indicates that nontuberculous mycobacterial infections should be considered when conventional microbiological assays fail to identify the infecting agent in suspected osteomyelitis following open fracture. A combination of radical debridement, including removal of infected bone, and prolonged antimicrobial therapy are required to eradicate the infection completely.
  3 5,836 244
CORRESPONDENCE
Spectrum of zygomycoses in north India: An institutional experience
S Mohapatra, M Jain, I Xess
July-September 2010, 28(3):262-262
DOI:10.4103/0255-0857.66471  PMID:20644321
  2 2,115 201
Chryseobacterium meningosepticum cellulitis and sepsis in an adult female with pemphigus vulgaris
S Sood, V Nerurkar, S Malvankar
July-September 2010, 28(3):268-269
DOI:10.4103/0255-0857.66481  PMID:20644326
  2 3,104 143
SPECIAL ARTICLES
The incredible journey of mankind: Helicobacter pylori as the narrator
P Desikan
July-September 2010, 28(3):193-195
DOI:10.4103/0255-0857.66466  PMID:20644304
Over the past decade, sequence differences between microbes from various geographical areas have been studied with the intent to interpret population movements of their hosts. An organism that is a reliable storehouse of such data, by virtue of its long association with its human host, is Helicobacter pylori. Functional and comparative analyses of its genome provide fascinating insights into human behaviour in the ancient past.
  2 6,119 584
BRIEF COMMUNICATIONS
Comparison of an in-house latex agglutination test with IgM ELISA and MAT in the diagnosis of leptospirosis
S Shekatkar, NS Acharya, BN Harish, SC Parija
July-September 2010, 28(3):238-240
DOI:10.4103/0255-0857.66484  PMID:20644313
The laboratory diagnosis of leptospirosis is fraught with several problems. Isolation of Leptospira by culture has a low sensitivity and the microscopic agglutination test (MAT) is time consuming To overcome these problems, a rapid latex agglutination test (LAT) has been standardized for the detection of antileptospiral antibodies in serum samples from suspected cases of leptospirosis. We compared the efficiency of the LAT to a commercially available IgM ELISA and MAT. A total of 150 serum samples were tested by LAT, IgM ELISA, and MAT. The positivity was 26.7%, 26% and 24% respectively. The sensitivity and specificity of LAT as compared to MAT was 90.62 and 91.96% respectively. Even though LAT and ELISA showed similar results, its rapidity and simplicity made latex agglutination test more suitable as a rapid screening test.
  1 5,313 387
CASE REPORTS
Early onset primary pulmonary cryptococcosis in a renal transplant patient
B Tarai, V Kher, P Kotru, A Sabhikhi, P Barman, A Rattan
July-September 2010, 28(3):250-252
DOI:10.4103/0255-0857.66489  PMID:20644317
We report a case of primary pulmonary cryptococcosis in a post-renal transplant patient. A 65-year-old male renal transplant patient was admitted to the hospital with a low grade fever of 1 month, radiologically mimicking tuberculosis (TB). Broncho-alveolar fluid (BAL) shows capsulated yeast, and Cryptococcus neoformans was grown on culture supported by cytology and histopathological examination. Cryptococcal antigen was positive (32-fold) in serum and was negative in cerebrospinal fluid (CSF). The patient was given amphotericin B and 5-flucytosine and clinical improvement was seen on a weekly follow up. The serum cryptococcal antigen test might contribute to the early detection and treatment of pulmonary cryptococcosis. The results of antifungal susceptibility were aid in selecting the drug of choice for treatment.
  1 3,245 171
Fungal endophthalmitis caused by Paecilomyces variotii, in an immunocompetent patient, following intraocular lens implantation
KB Anita, N Fernandez, R Rao
July-September 2010, 28(3):253-254
DOI:10.4103/0255-0857.66491  PMID:20644318
We report the case of a 70-year-old man who was admitted for anterior endophthalmitis following an intraocular lens implantation. He had developed a fluffy growth resembling a fungal mass on the iris of the right eye. The mass was removed and sent for fungal studies to our department. Direct microscopy revealed hyphae. Further studies helped identify the fungus to belong to genus Paecilomyces. This is a rare case of fungal endophthalmitis caused by Paecilomyces variotii in an immunocompetent person.
  1 3,825 148
A variant epidemic methicillin resistant Staphylococcus aureus-15 cavernous sinus thrombosis and meningitis: A rare occurrence with unusual presentation
HB Veena Kumari, D Nagaraja, S Nagarathna, GB Kulkarni, CS Praveen, S Nadig, G Arakere
July-September 2010, 28(3):255-257
DOI:10.4103/0255-0857.66492  PMID:20644319
Septic cavernous sinus thrombosis (CST) is an uncommon clinical syndrome. Although Staphylococcus aureus (S aureus) is the most common bacterial pathogen causing CST, it is infrequent as a cause of meningitis. We report the first case of CST and meningitis from Bengaluru, Karnataka, caused by community-acquired epidemic methicillin resistant Staphylococcus aureus-15 (EMRSA-15), in a previously healthy individual without known risk factors; the patient recovered following treatment with vancomycin. The isolate was genotyped as belonging to staphylococcal cassette chromosome mec type IV and sequence type 22 and carried the panton-valentine leucocidin gene. It is the first Indian EMRSA-15 disease isolate from a case of meningitis. EMRSA-15 has been a major problem in hospitals in UK and it is a cause for great concern in Indian hospitals too.
  1 4,678 264
CORRESPONDENCE
High prevalence of co-expression of newer β-lactamases (ESBLs, Amp-C-β-lactamases, and metallo-β-lactamases) in gram-negative bacilli
SS Chatterjee, R Karmacharya, SK Madhup, V Gautam, A Das, P Ray
July-September 2010, 28(3):267-268
DOI:10.4103/0255-0857.66479  PMID:20644325
  1 3,804 572
RNA positivity rates among anti-HCV reactive blood donors in Sri Lanka: A preliminary study
A Manamperi, P Nugawela, NS Gunawardene, Wimaladharma Abeyewickreme, J de Silva
July-September 2010, 28(3):264-265
DOI:10.4103/0255-0857.66475  PMID:20644323
  1 5,430 148
ORIGINAL ARTICLES
Comparison of Helicobacter pylori and Escherichia coli in induction of TNF-α mRNA from human peripheral blood mononuclear cells
E Ahmadzadeh, H Zarkesh-Esfahani, R Roghanian, F Navab Akbar
July-September 2010, 28(3):233-237
DOI:10.4103/0255-0857.66482  PMID:20644312
Purpose: To investigate the difference between the abilities of Helicobacter pylori and Escherichia coli to induce expression of TNF-α in human peripheral blood mononuclear cells (PBMC). Materials and Methods: H pylori was isolated from gastric biopsy specimens. The mononuclear cells were isolated from human blood, cultured, and treated with either intact or sonicated E coli or H pylori, and mRNA expression for TNF-α was detected using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Results: TNF-α mRNA expression levels were significantly higher in PBMCs stimulated with E coli compared to those stimulated with H pylori at the same number and identical conditions (P < .001). The results also suggest that sonicated bacteria were significantly (P < .001) less stimulatory for PBMCs than intact bacteria for both E coli and H pylori. Conclusions: The ability of different H pylori strains isolated from biopsy samples to stimulate TNF-α from PBMCs was significantly lower than that of E coli. Sonicated bacteria, as compared to intact bacteria, was a very poor inducer of TNF-α mRNA expression, suggesting that the conformation of lipopolysaccharides (LPS) on the outer leaflet of the outer membrane is not totally conserved in sonicated bacteria.
  1 12,620 214
Kinetics of chemokine secretion in human macrophages infected with various strains of Mycobacterium tuberculosis
P Rajavelu, SD Das
July-September 2010, 28(3):201-206
DOI:10.4103/0255-0857.66470  PMID:20644306
Background and Purpose: It has been shown that chemokine secretion upon infection with Mycobacterium tuberculosis is influenced by the virulence of the strain, and it is suggested that virulence-associated differences in chemokine secretion contribute to the failure in containing the infection due to poor granuloma formation. Materials and Methods: In this study, we used prevalent M tuberculosis clinical strains (S7 and S10) to study the chemokine secretion profile in infected THP-1 cells and monocyte-derived macrophages (MDM) and compared this with the chemokine secretion induced by laboratory strains. Results: This study showed that comparatively lower levels of IP-10 were induced by clinical strains than by laboratory strains in both differentiated THP-1 and MDMs. The secretion of MIP-1α was also depressed but only in the THP-1 cells infected with clinical strains. This depressed chemokine secretion may hinder the movement of Th-1 cells from the periphery into the infection foci to control the infection. Correlation between IP-10 and IL-12p40 showed a negative relationship in control MDMs, while there was a positive correlation in all the infected strains, indicating their cooperative role in attracting and activating Th1 cells for a protective immune response at the site. This relationship was strain dependent, with avirulent H37Ra showing higher correlation, followed by the clinical strains and the virulent H37Rv. A positive correlation of IP-10 with IFN-γ (S7 and H37Ra) and with IL-10 (H37Ra and H37Rv) suggested a definitive interplay of these molecules in infection. Conclusions: The chemokines secretion by infected THP-1 cells and MDMs was strain dependant and the lower induction by the clinical strains may indicate that the clinical strains maintain a quiescent nature to mislead the host immune system for their benefit.
  1 4,174 301
KatG protein: A novel marker for differential diagnosis of Myobacterium avium complex infection
K Gupta, I Verma, G Khuller, R Mahajan
July-September 2010, 28(3):221-226
DOI:10.4103/0255-0857.66478  PMID:20644310
Purpose: Biochemical or nucleic acid based diagnostic techniques for MAC infection are unsatisfactory. This study aims to identify and evaluate M. avium secretory protein(s) of diagnostic potential, so as to develop a rapid and simple method for diagnosis of MAC infection. Material and Methods: Initially, a specific protein band of ~80-85 kDa was recognised by differential immunoblotting; which was subjected to anion exchange column chromatography for purification of proteins. After fractionisation using SDS-PAGE and electroelution, blast search was carried out. Further immunoreactivity studies were done with M. avium and Mtb infected mice sera. Clinical utilisation of separated protein was evaluated by conducting indirect ELISA with serum samples from mycobacterial infected patients. Results: A specific 81.6 kDa protein, shown to be catalase-peroxidase protein (KatG) by blast search was separated. Immunoreactivity studies of purified KatG proteins with mice sera confirmed it to be specific for M. avium infection. Indirect ELISA with patient samples further confirmed it to be M. avium infection specific. Conclusion: KatG protein is specifically recognised by MAC patients and can be used as a marker for simple and rapid ELISA based tests for differential diagnosis of M. avium infection.
  1 4,067 303
BOOK REVIEW
Emergencies in Infectious Diseases: From head to toe
S Badrinath
July-September 2010, 28(3):271-271
  - 1,868 216
BRIEF COMMUNICATIONS
A cross-sectional hospital-based study of autoantibody profile and clinical manifestations of systemic lupus erythematosus in south Indian patients
K Kosaraju, S Shenoy, U Suchithra
July-September 2010, 28(3):245-247
DOI:10.4103/0255-0857.66487  PMID:20644315
Our study was aimed to analyze clinical manifestations, autoantibodies and other serological abnormalities in South Indian patients with systemic lupus erythematosus. Clinical history and findings on systemic examination were noted. Antinuclear antibodies (ANA), antibodies to double-stranded DNA (dsDNA) were detected by immunofluorescence and ANA profile by Immunoblotting. Arthritis was most common followed by fever and skin rash. Clinical manifestations vary according to geographical location of the patient. ANA was positive in 64.28% and anti-dsDNA in 89.36% of patients. All patients with lupus nephritis were positive for dsDNA. Detection of antibodies to dsDNA, RNP and anti-Smith (Sm) are of diagnostic and prognostic importance.
  - 4,294 288
CORRESPONDENCE
Trend of syphilis in Pakistan, 1991-2008
E Khan, BI Memon, A Ayaz, F Malik, SF Shamim, FS Polani, A Zafar
July-September 2010, 28(3):263-264
DOI:10.4103/0255-0857.66473  PMID:20644322
  - 3,492 112
Recurrent infections with Aspergillus require exclusion of defects in the phagocyte NADPH oxidase pathway
S Khan
July-September 2010, 28(3):269-270
DOI:10.4103/0255-0857.66483  PMID:20644327
  - 2,185 96
Comment: Infection control with limited resources: Why and how to make it possible?
S Joshi
July-September 2010, 28(3):270-270
DOI:10.4103/0255-0857.66485  PMID:20644328
  - 2,081 215
ORIGINAL ARTICLES
Development of polyclonal antisera to clone enterovirus 71 cellular receptor
AB Sioofy-Khojine
July-September 2010, 28(3):196-200
DOI:10.4103/0255-0857.66469  PMID:20644305
Purpose: Enterovirus 71 (ENV71) is a member of Picornaviridae family and was shown to be of public health concern in the Far East because of the notorious outbreaks it caused, with novel clinical features in the affected patients. In this study we assessed the use of virus capsid protein VP1 in viral receptor research. Material and Methods: The capsid protein (VP1) was cloned, expressed in a prokaryotic system, and purified for immunisation of rabbits. The immunisation was carried out according to the UK Home Office regulations. The polyclonal antisera were collected and tested for reactivity against recombinant and native VP1 of ENV71. Results: Both antisera were reactive against native and partially/fully denatured viral particles. Conclusion: The antisera are functional in receptor studies.
  - 3,528 244
RESEARCH SNIPPETS
Research snippets from the medical world
P Desikan
July-September 2010, 28(3):272-273
DOI:10.4103/0255-0857.66467  
  - 1,795 146

2004 - Indian Journal of Medical Microbiology
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Online since April 2001, new site since 1st August '04