PMID- 15364272
OWN - NLM
STAT- MEDLINE
DCOM- 20041210
LR  - 20061115
IS  - 1386-6532 (Print)
IS  - 1386-6532 (Linking)
VI  - 31
IP  - 2
DP  - 2004 Oct
TI  - Detection of hepatitis C virus by a user-developed reverse transcriptase-PCR and 
      use of amplification products for subsequent genotyping.
PG  - 148-52
AB  - BACKGROUND: Detection, quantitation and genotyping of hepatitis C virus (HCV) are 
      important in selecting appropriate therapy. Current commercially available HCV 
      genotyping kits, including sequencing-based TRUGENE HCV 5'NC and 
      hybridization-based INNO-LiPA HCV II assays, rely on amplification products 
      (amplicons) generated by HCV reverse-transcriptase (RT)-PCR methods such as the 
      Roche AMPLICOR HCV test. METHODS: We developed a one-step RT-PCR assay to amplify 
      and detect HCV RNA, and the resulting amplicons were used for HCV genotyping 
      (TRUGENE). A total of 142 clinical samples were used to compare results from the 
      RT-PCR/TRUGENE assay and those generated by the COBAS AMPLICOR and INNO-LiPA 
      tests. RESULTS: Eighty-seven of 108 plasma specimens which were positive by 
      AMPLICOR were also positive by the user-developed RT-PCR, giving a sensitivity of 
      100.0%. The RT-PCR detected 2 of 21 AMPLICOR-negative specimens and none of 34 
      HCV-EIA-negative serum specimens, giving a specificity of 96.4%. The 87 amplicons 
      from the RT-PCR yielded HCV genotypes. HCV genotype results from both TRUGENE and 
      INNO-LiPA were all in agreement. The TRUGENE and INNO-LiPA assays identified 69 
      (79.3%) and 47 (54.0%) specimens, respectively at the subtype level. HCV subtype 
      information agreed by both assays in 34 of 36 (99.4%) specimens. One specimen 
      with HCV genotypes 2 and 4 by INNO-LiPA was classified as a single genotype 2 by 
      TRUGENE. CONCLUSION: Our data showed that the user-developed RT-PCR has 
      comparable sensitivity and specificity for the detection of HCV in clinical 
      specimens. The amplicons generated by the RT-PCR can be used for HCV genotyping 
      by the sequencing-based TRUGENE assay.
FAU - Tang, Yi-Wei
AU  - Tang YW
AD  - Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 
      37232, USA. yiwei.tang@vanderbilt.edu
FAU - Li, Haijing
AU  - Li H
FAU - Roberto, Ann
AU  - Roberto A
FAU - Warner, Diane
AU  - Warner D
FAU - Yen-Lieberman, Belinda
AU  - Yen-Lieberman B
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - Netherlands
TA  - J Clin Virol
JT  - Journal of clinical virology : the official publication of the Pan American 
      Society for Clinical Virology
JID - 9815671
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Base Sequence
MH  - DNA, Viral/genetics
MH  - Genotype
MH  - Hepacivirus/*genetics/*isolation & purification
MH  - Hepatitis C/diagnosis/virology
MH  - Humans
MH  - Nucleic Acid Amplification Techniques
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Virology/methods
EDAT- 2004/09/15 05:00
MHDA- 2004/12/16 09:00
CRDT- 2004/09/15 05:00
PHST- 2004/02/06 00:00 [revised]
PHST- 2004/02/12 00:00 [accepted]
PHST- 2004/09/15 05:00 [pubmed]
PHST- 2004/12/16 09:00 [medline]
PHST- 2004/09/15 05:00 [entrez]
AID - S1386653204000484 [pii]
AID - 10.1016/j.jcv.2004.02.010 [doi]
PST - ppublish
SO  - J Clin Virol. 2004 Oct;31(2):148-52. doi: 10.1016/j.jcv.2004.02.010.