Evaluation of three molecular biology-based assays for the detection of GB virus C/hepatitis G virus in clinical specimens.
OBJECTIVE
The present study was performed to evaluate the reliability of three reverse transcription-polymerase chain reaction (RT-PCR) assays, one commercial and two 'homebrew', for GB virus C (GBV-C)/hepatitis G virus (HGV) RNA detection in clinical specimens. We, therefore, investigated the virus prevalence with the method that gave us the best performances.


METHODS
The commercial assay amplified sequences from the viral 5'-untranslated region (5'UTR) and non-structural 3 (NS3) region. The non-commercial assays 1 and 2 were based on different primers for the 5'UTR consensus sequence.


RESULTS
The percentage of overall concordance by the three methods was 91.7%, raising to 93.0% when only the two non-commercial methods were compared. Assay 1 showed low sensitivity (57.1% vs. the commercial assay, 58.8% vs. assay 2), with 100% specificity. The commercial assay gave 18 of 54 (33.3%) 'false-negative' results, concordantly negative by the other assays. The prevalence of GBV-C/HGV RNA among the HIV+ patients was 27.0 and 32.6% in HIV/HCV co-infected patients.


CONCLUSIONS
These data suggest that assay 2 has higher reliability as compared to the other two methods and may be used for an accurate GBV-C/HGV RNA detection in clinical and epidemiological studies.
Bortolin
Maria Teresa
MT
Microbiology-Immunology and Virology Department, Centro di Riferimento Oncologico, IRCCS, Aviano, Italy. micro@cro.it
Zanussi
Stefania
S
Tedeschi
Rosamaria
R
Pratesi
Chiara
C
D'Andrea
Monica
M
Bidoli
Ettore
E
di Gennaro
Giampiero
G
De Paoli
Paolo
P
D020121:5' Untranslated Regions; D029407:GB virus C / Q000235:genetics / Q000302:isolation & purification*; D015658:HIV Infections / Q000150:complications; D006525:Hepatitis, Viral, Human / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D012367:RNA, Viral / Q000032:analysis / Q000097:blood; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000235:genetics
D020121:5' Untranslated Regions; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D017361:Viral Nonstructural Proteins