Real-time fluorescent quantitative PCR primer pair and detection kit for detecting hepatospora eriocheir and application of detection kit

Abstract

The invention discloses a real-time fluorescent quantitative PCR primer pair for detecting hepatospora eriocheir. Sequences of the primer pair are shown as SEQ ID NO:1 and SEQ ID NO:2. The invention further discloses a real-time fluorescent quantitative PCR detection kit and application thereof. The primer pair, the kit and a detection method, provided by the invention, have the following advantages in detection of the hepatospora eriocheir: no cross-reaction occurs with other pathogenic microsporidia, and the specificity is strong; variation coefficients of repeatability test results within agroup and between groups are all less than 2%, and the repeatability is good; and the amplification efficiency is 98.3%, and the PCR primer pair can detect 2.87*101 copies / [mu]L at lowest, which is 100 times that of an ordinary PCR. The detection method established by the invention is higher in amplification efficiency and detection sensitivity; and a whole experimental process only takes 2 hoursfrom extraction of DNA from the to-be-detected hepatospora eriocheir to a result analysis, and the detection efficiency is greatly improved.

Description

technical field [0001] The invention relates to the field of microbial molecular detection, in particular to a pair of real-time fluorescent PCR primers, a detection kit and an application for detecting Hepatocystoma mitten crab. Background technique [0002] Chinese mitten crab (Eriocheir sinensis), also known as river crab and hairy crab, is deeply loved by consumers because of its delicious taste and rich nutrition. It is the main economic crab species cultivated in my country. However, with the development of intensive farming, various diseases of river crabs occur frequently, among which "water slug" is one of the diseases that have caused the most serious damage to river crabs in recent years. The symptoms caused by it are mainly manifested as hepatopancreas The color changes, from the healthy yellow hepatopancreas to lighter to milky white and grayish black, pleural effusion, hepatopancreas and muscle atrophy or even disappear, and other symptoms have brought huge eco...

AI Technical Summary

Problems solved by technology

Although these methods can achieve the purpose of detecting Hepatocystis in mitten crab, there are still some deficiencies, such as high missed diagnosis rate, prone to false positives, relatively low sensitivity, cumbersome operation, and long time-consuming. For example, the method established by Ding Zhengfeng The real-time fluorescence quantitative PCR detection method of Hepatosoma mitteni has only 79.1% amplification efficiency, and the lower detection limit of sensitivity can only reach 200 copies / μL, which is not suitable for the detection of Hepatocytes of mitten crab in low-concentration samples

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