Sensitive and specific detection of hepatitis C virus RNA with nested polymerase chain reaction and digoxigenin labeled cDNA probe.

一彰, 茶山; 博光, 熊田; 聡, 斉藤; 康司, 荒瀬; 健次, 池田; 洋子, 酒井; 豊海, 松本; 万利子, 小林; 傳, 森永

doi:10.2957/kanzo.32.136

Cited by 4 publications

(2 citation statements)

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“…Nested PCR is an extremely sensitive method that can detect even a single copy of a viral sequence (11)(12)(13). The disappearance of HCV RNA in serum as detected by this method during therapy suggests that IFN is effective against chronic hepatitis C in some patients, presumably because of its antiviral effects.…”

Section: Discussionmentioning

confidence: 99%

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Effect of interferon administration on serum hepatitis C virus RNA in patients with chronic hepatitis C

et al. 1991

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Hepatitis C virus RNA as detected by reverse transcription and nested polymerase chain reaction was monitored in 16 patients with chronic hepatitis C treated with interferon. Hepatitis C virus RNA became undetectable after 4 to 8 wk of interferon administration in 13 of the 16 patients. During 6 mo of follow-up, 5 of the 13 patients who became negative for hepatitis C virus RNA after interferon administration remained negative, and all five continued to have normal ALT levels. Repeat liver biopsy in these five patients revealed histological improvement. Antibody to hepatitis C virus, which was initially positive in all treated patients, fell to undetectable levels in three of the five patients. In contrast, aminotransferase levels rose again in all eight patients who had become hepatitis C virus RNA negative but had again exhibited hepatitis C virus RNA after completion of therapy. In 16 untreated patients, hepatitis C virus RNA remained detectable. These results indicate that detection of hepatitis C virus RNA may be useful as a marker of viral replication in chronic hepatitis C; they also suggest that interferon should again be administered to patients who become hepatitis C virus RNA negative on treatment but again exhibit this marker of viral replication when treatment is stopped.

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Section: Discussionmentioning

confidence: 99%

“…With the advent of the polymerase chain reaction (PCR) (101, it has become possible to detect small amounts of HCV RNA sequences in serum. We have detected HCV RNA in serum using a sensitive nested-PCR method (11)(12)(13) and show here that the disappearance and reappearance of HCV RNA is closely related to the outcome of disease.…”

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confidence: 87%

Effect of interferon administration on serum hepatitis C virus RNA in patients with chronic hepatitis C

et al. 1991

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Usefulness of HCV-RNA counts by the method of HCV-bDNA probe in interferon retreatment for patients with HCV-RNA positive chronic hepatitis C

Arase¹,

Kumada²,

Chayama³

et al. 1995

International Hepatology Communications

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Quantitative analysis of hepatitis C virus RNA by competitive nested polymerase chain reaction

Chayama

Tsubota

Arase

et al. 1993

J of Gastro and Hepatol

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A method that allows the quantitation of hepatitis C virus (HCV) RNA is described. The RNA was extracted from serum samples and reverse transcribed. Target cDNA was then co‐amplified by nested polymerase chain reaction with a known amount of competitive template at various concentrations. Since this internal control DNA uses the same primers as those of the target and is distinguishable from the target cDNA after amplification by size, the initial concentration of the target could be estimated by comparing the intensity of the two bands of amplified DNA fragments. That is, if the starting amount of the cDNA and the competitive template are equal, the intensity of the two bands should also be equal. Using this method the amount of HCV RNA in serum samples obtained from 85 patients with chronic hepatitis type C was determined. There was as much as a 100 000‐fold difference in the levels of HCV RNA from patient to patient. A rapid decrease of HCV RNA in a patient treated with interferon is also described.

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Sensitive and specific detection of hepatitis C virus RNA with nested polymerase chain reaction and digoxigenin labeled cDNA probe.

Cited by 4 publications

References 10 publications

Effect of interferon administration on serum hepatitis C virus RNA in patients with chronic hepatitis C

Effect of interferon administration on serum hepatitis C virus RNA in patients with chronic hepatitis C

Usefulness of HCV-RNA counts by the method of HCV-bDNA probe in interferon retreatment for patients with HCV-RNA positive chronic hepatitis C

Quantitative analysis of hepatitis C virus RNA by competitive nested polymerase chain reaction

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