| ORIGINAL ARTICLE |
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| Year : 2020 | Volume
: 38
| Issue : 3 | Page : 430-439 |
Two novel genomic DNA sequences as common diagnostic targets to detect Cryptosporidium hominis and Cryptosporidium parvum: Development of quantitative polymerase chain reaction assays, and clinical evaluation
Arpit Kumar Shrivastava1, Swagatika Panda2, Subrat Kumar3, Priyadarshi Soumyaranjan Sahu4
1 Infection Biology Laboratory, School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha; Department of Microbiology, Virus Research and Diagnostic Laboratory, Atal Bihari Vajpayee Government Medical College, Vidisha, Madhya Pradesh, India
2 Infection Biology Laboratory, School of Biotechnology, KIIT Deemed to be University; Department of Infectious Disease Biology, Institute of Life Sciences, Bhubaneswar, Odisha, India
3 Infection Biology Laboratory, School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha, India
4 Infection Biology Laboratory, School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha, India; Department of Microbiology and Immunology, Medical University of the Americas (R3 Education Inc), MA, USA
Correspondence Address:
Dr. Priyadarshi Soumyaranjan Sahu
Department of Microbiology and Immunology, Medical University of the Americas (R3 Education Inc.), 27 Jackson Road # 300, Devens, MA 01434
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/ijmm.IJMM_20_114
Introduction: Cryptosporidium is an intestinal parasite responsible for gastroenteritis. Conventional diagnosis of Cryptosporidium is made by microscopy. The most frequent molecular detection method for this parasite is polymerase chain reaction (PCR). The objective of the present study was to identify the novel DNA targets and development of PCR-based assays for the specific detection of two major human infecting species Cryptosporidium parvum and Cryptosporidium hominis. Methodology: Sensitive and specific SYBR green quantitative PCR (qPCR) and TaqMan qPCR assays were developed and validated at both diagnostic and analytical level using the new identified targets TU502HP-1 and TU502HP-2. Results: Assay validation results showed that the newly developed real-time PCR assays are 100% specific with a reliable limit of detection. Overall repeatability and reproducibility of these assays showed good quality results over intra- and inter-laboratory analysis. Conclusion: Novel target-based qPCR assays can be rapid an efficient tool for simultaneous detection of a C. parvum and C. hominis. These genes could also be utilized for the development of innovative DNA-based Point-of-Care test development.
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