Diagnosis reagent for non-A non-G hepatitis and its detection method

Abstract

The present invention features the combined use of PCR technology and microwell plate nucleic acid hybrid enzyme coloring technology in the clinical diagnosis of non-A to non-G hepatitis. In the diagnosis reagent and detection method, single PCR, rather than twice PCR amplification, is adopted, and biotin-avidin is applied to coat the PCR product onto microwell plate and to make it hybridized with fluorescein-labeled probe, so that the specific enzyme label and fluorescein antibody produce coloring reaction. The present invention eliminates the influence of non-specific amplification product, avoid false positivity, and obtains reliable result. The present invention has high sensitivity, high specificity and no ethidium bromide pollution, and is fast, accurate and repeatable.

Description

technical field [0001] The invention relates to the technical field of disease diagnosis, in particular to a non-A-G hepatitis diagnostic reagent and a detection method thereof. Background technique [0002] In the etiological diagnosis of hepatitis, about 10%-20% of acute and chronic post-transfusion hepatitis, sporadic hepatitis, liver cirrhosis and fulminant hepatitis cannot detect their virological signs with existing detection methods, suggesting that there may be non-viral markers. AG virus. In 1997, Japan isolated a new DNA virus called transfusion-transmitted virus (transfusion-transmitted virus, TTV) in the serum of patients with non-AG hepatitis after blood transfusion. It is very likely to become the eighth hepatitis virus. Studies have shown that TTV has a global distribution and is a single-stranded circular DNA virus without a cell membrane. It is related to abnormally elevated transaminases and abnormal liver function in patients with unexplained non-A-G hepa...

AI Technical Summary

Problems solved by technology

[0003] Because the serum TTV DNA titer of TTV infected patients is 50-5000 copies / ml, compared with some other viruses transmitted through blood (such as HCV RNA titer is 10 3 -10 copies / ml) is low, therefore, it is not yet possible to use serum immunological diagnostic methods to detect TTV, and the use of polymerase chain reaction (PCR) has become the main means of diagnosing TTV infection

Although PCR technology has high specificity and sensitivity, the detection of TTV requires two PCR amplifications. The operation is cumbersome and the technical requirements are high. It is easy to cause cross-contamination and false positives, thus affecting the judgment of the results.