Clinical usefulness of a new hepatitis C virus RNA extraction method using specific capture probe and magnetic particle in hemodialysis patients.
Hemodialysis patients are a high-risk group for hepatitis C virus (HCV) infection. Assessment of HCV infection using HCV-RNA assay among dialysis patients is important for the issue of safety and environmental protection. However, polymerase chain reaction (PCR)-based methods are unsuitable for analyzing samples from dialysis patients because the conventional centrifugal extraction method fails to eliminate heparin, a potent inhibitor of PCR. In this study, we evaluated the usefulness of a HCV-RNA extraction method using probes and magnetic particles for hemodialysis patients in comparison with the centrifugal method. The study population consisted of 17 HCV antibody-positive patients undergoing hemodialysis. These 17 patients consisted of 12 HCV carrier patients and five patients with past HCV infection. One hundred and two samples from these patients were measured using the centrifugal and magnetic methods. Moreover, we prepared five standards that included theoretically 5 KIU/mL of HCV. One was made from non-HD patient's serum and the other four were from hemodialysis patients' serum. These standards were measured using the two methods. False-negative results were not observed with the magnetic method, but were observed in five out of 102 samples with the centrifugal method. Studies using standard samples revealed that accurate HCV-RNA measurement is achieved using the magnetic method. In conclusion, the present study showed that this magnetic extraction method is a highly reproducible and reliable assay to obtain correct information about the presence of the infective virus itself in the hemodialysis setting. Precise identification of HCV-RNA using this specific method is considered to be useful in preventing HCV infection in hemodialysis units.
Department of Nephrology, Akebono Clinic, Kumamoto, Japan. tanaka@matusita-kai.or.jp
Motoko
M
Tanaka
Shigetoshi
S
Fujiyama
Motohiko
M
Tanaka
Kazuko
K
Itoh
Kazunori
K
Matsushita
Kazutaka
K
Matsushita
Kazuhiro
K
Matsuyama
Hirokazu
H
Kakuda
Kimio
K
Tomita
D000368:Aged; D019921:Biotinylation; D002498:Centrifugation; D005260:Female; D016174:Hepacivirus / Q000276:immunology; D006526:Hepatitis C / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D008280:Magnetics; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D006435:Renal Dialysis; D015203:Reproducibility of Results; D019562:Viral Load
D012367:RNA, Viral