Hepatitis B virus YMDD motif mutation detection specific primer and liquid phase chip and method thereof

Abstract

The invention discloses a hepatitis B virus YMDD motif mutation detection specific primer and liquid phase chip and a method thereof. The liquid phase chip mainly comprises an ASPE primer, a microsphere coated with an anti-tag sequence and a primer used for amplifying target sequences having mutation sites of rtL801 / V, rtV173L, rtL180M, rtA181V / T, rtM204V / I / S and / or rtN 236T. The detection liquid phase chip and the detection method have high detection sensitivity, high signal-to-noise ratio and more accurate and reliable detection results. Moreover, detecting 6 mutation sites can be completed in the same reaction, i.e., the operation is convenient, and many uncertain factors existing in many times of operational processes are avoided, thereby greatly improving the detection accuracy.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, and in particular relates to specific primers for detecting mutations of viral YMDD motifs related to drug resistance of hepatitis B nucleoside analogs, a liquid phase chip and a detection method thereof. technical background [0002] Hepatitis B is a globally widespread infectious disease with geographical distribution in many countries. There are more than 2 billion people in the world who have been infected with hepatitis B virus (hepatitis B virus, HBV), of which 350-400 million are chronic carriers, and there are 120 million people in China. Chronic hepatitis B can cause liver cirrhosis, liver failure, liver cancer and other serious liver diseases, and about 1 million people die every year in the world. And China is the hardest-hit area of ​​chronic hepatitis B, and more than 8% of the population are chronic carriers, and about 300,000 people die of li...

AI Technical Summary

Problems solved by technology

Although direct sequencing has always been considered the gold standard for detecting gene mutations, it is time-consuming and laborious, does not meet the high-throughput requirements of clinical testing, and its sensitivity is not high

RFLP also has complicated operation steps, and the test results are affected by many aspects, so it cannot meet the needs of large-scale, rapid and accurate clinical testing.

Although solid-phase gene chips can achieve high-throughput detection, they are expensive, have low sensitivity, and poor reproducibility of test results

The above techniques are less capable of detecting inferior mutant virus strains in mixed infections, so it is impossible to detect YMDD mutant strains of hepatitis B virus in a timely and early manner clinically.

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