Evaluation of the Efficiency of HCV RT-PCR Using Narrowly Spaced Primers

Jang, Sook Jin; Cho, Song Sig; Kang, Jeong Suk; Park, Geon; Moon, Dae Soo; Park, Young Jin

Abstract

BACKGROUND
It is important to choose effective primers to increase the sensitivity of RT PCR for HCV. It was reported that the use of narrowly spaced primers (NSP) resulted in increased sensitivity of HCV PCR because NSP is more effective than widely spaced primers (WSP) in RT or PCR reaction. It will be useful to compare the merits and demerits of PCR using NSP (NSP-PCR), PCR using WSP (WSP-PCR) and anti-HCV EIA.
METHODS
We performed NSP-PCR, WSP-PCR, anti-HCV EIA tests and determined sensitivity and specificity of each method with 36 sera from patients with hepatitis C and 14 sera from patients with diseases other than hepatitis C.
RESULTS
The sensitivity and specificity for WSP-PCR were 80.6% and 100%, those for NSP-PCR were 86.1% and 78.6%, and those for anti-HCV EIA were 94.4% and 100%. The concordance rate for anti-HCV EIA and WSP-PCR was 82% and that of anti-HCV EIA and NSP-PCR was 80%. The detection limit of WSP-PCR was up to 10(-1) dilution and that of NSP-PCR was up to 10(-2) dilution. The positive bands of WSP-PCR were strong and large. But those of NSP-PCR were weak and small, so it was not easy to differentiate them from primer dimers sometimes.
CONCLUSIONS
WSP-PCR seemed to be better than NSP-PCR for clinical laboratory tests. Anti-HCV EIA appears to be a good screening test for liver disease because of the high sensitivity and specificity.