PCR Detection of Group A Bovine Rotaviruses in Feces
| Autor: | Geoffrey Y. Akita, Anthony E. Castro, Bennie Osburn, Jarasvech Chinsangaram |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Rotavirus
0301 basic medicine 040301 veterinary sciences Molecular Sequence Data 030106 microbiology Pcr cloning Cattle Diseases Enzyme-Linked Immunosorbent Assay Biology Kidney medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Group A Rotavirus Infections Cell Line law.invention 0403 veterinary science Feces 03 medical and health sciences chemistry.chemical_compound law medicine Animals Polymerase chain reaction DNA Primers RNA Double-Stranded Chemiluminescence Base Sequence General Veterinary RNA 04 agricultural and veterinary sciences Macaca mulatta Virology Molecular biology chemistry RNA Viral Agarose Cattle |
| Zdroj: | Journal of Veterinary Diagnostic Investigation. 5:516-521 |
| ISSN: | 1943-4936 1040-6387 |
| DOI: | 10.1177/104063879300500403 |
| Popis: | A polymerase chain reaction (PCR) protocol has been developed for identification of bovine group A rotavirus infection in feces. Primers (20mers) complementary to 3′ ends of double-stranded RNA genome segment 6 of bovine rotavirus NCDV strain were synthesized and used in PCR. Bovine rotavirus RNA from infected cell culture was employed to optimize the PCR protocol. Rotavirus-negative fecal samples were spiked with known quantities of bovine rotavirus, and the sensitivity of the PCR assay was determined. Fecal samples were extracted with phenol and treated to eliminate unidentified PCR inhibitor(s) in feces, and PCR was performed. PCR products were either visualized on ethidium bromide-stained agarose gels or detected by chemiluminescent hybridization. The sensitivity of the assay was 6 × 104 viral particles/ml of feces with ethidium bromide-stained agarose gel visualization or 6 × 102 viral particles/ml of feces with chemiluminescent hybridization. The PCR assay was applied to 18 fecal specimens from clinical cases. All 16 clinical samples that were positive for rotavirus by enzyme-linked immunosorbent assay (ELISA) or by ELISA and electron microscopy (EM) were positive by PCR. The 2 samples that were rotavirus negative by ELISA or by ELISA and EM were also negative on PCR analysis. |
| Databáze: | OpenAIRE |
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