Evaluation of hepatitis delta virus RNA levels during interferon therapy by analysis of polymerase chain reaction products with a nonradioisotopic hybridization assay

✍ Scribed by Elisabetta Cariani; Antonella Ravaggi; Massimo Puoti; Giovanni Mantero; Alberto Albertini; Daniele Primi

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 592 KB
• Volume: 15
• Category: Article
• ISSN: 0270-9139
• DOI: 10.1002/hep.1840150422

✦ Synopsis

We developed a nonradioisotopic assay for detection of hepatitis delta virus RNA in serum by combining reverse transcription of RNA, polymerase chain reaction of the resultant complementary DNA and enzyme linked immunoassay detection of the polymerase chain reaction products using a monoclonal antibody specific for double-stranded DNA. This DNA enzyme immunoassay had a limit of detection of cloned hepatitis delta virus RNA similar to that of standard PCR followed by Southern-blot hybridization ( -10 copies/sample) and was lo3 to lo4 times more sensitive than direct dot-blot hybridization ( -lo5 copies/sample). Serial serum samples from six patients with chronic hepatitis delta virus infection undergoing interferon therapy were analyzed by reverse transcription-polymerase chain reaction followed by both standard hybridization and DNA enzyme immunoassay. The results of both methods were comparable, revealing disappearance of hepatitis delta virus RNA after 3 to 6 m o of therapy in three patients, two of whom had also a significant decrease in ALT activity. The DNA enzyme immunoassay test is therefore a potentially useful method for therapeutic monitoring in chronic hepatitis delta virus infection and may contribute to a wider application of polymerase chain reaction in clinical laboratories. (HEPATOLOGY 1992;15

685-689.)

Hepatitis delta virus (HDV) is a defective RNA virus that requires the presence of HBV to be infectious (1). HDV can cause severe acute or fulminant hepatitis and can lead to severe chronic hepatitis if a chronic HBV carrier becomes superinfected with HDV (2).

The diagnosis of delta hepatitis is usually made by