A SIMPLE SINGLE-TUBE PROCEDURE OF PCR ASSAY FOR THE DETECTION OF HEPATITIS-C VIRUS-RNA

Citation
Y. Hayashi et al., A SIMPLE SINGLE-TUBE PROCEDURE OF PCR ASSAY FOR THE DETECTION OF HEPATITIS-C VIRUS-RNA, Research in virology, 145(2), 1994, pp. 123-128
Citations number
15
Categorie Soggetti
Virology
Journal title
ISSN journal
09232516
Volume
145
Issue
2
Year of publication
1994
Pages
123 - 128
Database
ISI
SICI code
0923-2516(1994)145:2<123:ASSPOP>2.0.ZU;2-E
Abstract
We developed a simple single tube procedure using the PCR (polymerase chain reaction) for the detection of hepatitis C virus RNA. The entire reaction from RNA extraction to PCR occurred in one tube; this was ma de possible by the use of more than 1,000 mu g/ml of proteinase K for RNA extraction, instead of the acid-guanidinium thiocyanate-phenol-chl oroform method. All necessary reagents were added to the tube, and PCR products were not removed from the tube until the end of PCR, althoug h opening of the tube during the procedure could not be avoided. There fore, cross-contamination which might theoretically take place during transfer of products between tubes never occurred. This method detecte d about 1 chimpanzee infectious dose of HCV in H strain plasma.