Evaluation of different genomic regions of Rotavirus A for development of real time PCR
| Autor: | Shital G. Deore, Sujata S. Ranshing, Shobha D. Chitambar, Atul M. Walimbe, Madhuri S. Joshi |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Rotavirus Serial dilution Genotype viruses 030106 microbiology Genome Viral Biology Viral Nonstructural Proteins medicine.disease_cause Real-Time Polymerase Chain Reaction Sensitivity and Specificity Rotavirus Infections 03 medical and health sciences Feces Virology Human rotavirus medicine Humans Gene Antigens Viral Phylogeny virus diseases RNA Gastroenteritis 030104 developmental biology Real-time polymerase chain reaction Child Preschool RNA Viral Capsid Proteins Breast feeding |
| Zdroj: | Journal of virological methods. 266 |
| ISSN: | 1879-0984 |
| Popis: | The nucleotide alignment of all 11 genes of human Rotavirus A (RVA) strains revealed suitability of NSP2, NSP3 and VP6 genes for the development of real time PCR (qRT-PCR). Evaluation of qRT-PCR assays using known rotavirus ELISA positive and negative fecal specimens showed non-overlapping ranges of Mean ±3SD cycle threshold (Ct) values for NSP3 and VP6 based assays. Using serial dilutions of purified RVA, high sensitivity of VP6 qRT-PCR assay (1.95 × 10−5 pg/μL of RNA) was recorded as compared to NSP2 and NSP3 qRT-PCR assays (1.95 × 10-4 pg/μL of RNA). Further, evaluation of the VP6 qRT-PCR assay involving 266 fecal specimens and frequency polygon analysis of the data indicated cut-off value of 35 for Ct with high sensitivity (126/131, 96%) and specificity (12/12, 100%). This VP6 qRT-PCR assay will be a useful diagnostic tool to evaluate clinical presentations in rotaviral gastroenteritis under different conditions such as breast feeding and administration of rotavirus vaccines. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect