Real-time quantitative PCR (Polymerase Chain Reaction) method for detecting echinococcus multilocularis and echinococcus granulosus by types based on MGB (Minor Groove Binder) probe and detection kit

Abstract

The invention discloses a real-time quantitative PCR (Polymerase Chain Reaction) method for detecting echinococcus multilocularis and echinococcus granulosus by types based on an MGB (Minor Groove Binder) probe and a detection kit. The real-time quantitative PCR method comprises the following steps: designing primers, designing the MGB probe and selecting PCR amplification conditions; the detection kit contains an outer primer pair sequence of a PCR primer pair and an MGB type detection probe sequence. The method disclosed by the invention can be used for pertinently detecting samples containing the echinococcus multilocularis and / or the echinococcus granulosus by types and the accuracy is high; mitochondrion DNAs (Deoxyribonucleic Acid) of the echinococcus multilocularis and the echinococcus granulosus can be accurately detected by types; the sensitivity is high so that the echinococcus multilocularis and the echinococcus granulosus can be rapidly identified by types in a large batch; the operation is simple and convenient, and trace insect source mitochondrion DNAs in the samples can be amplified and the level of detecting by types is realized.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a real-time quantitative PCR technique typing detection method based on MGB probe typing detection. Background technique [0002] Real-time fluorescent quantitative PCR (Quantitative Real-time PCR) is a method of measuring the total amount of products after each polymerase chain reaction (PCR) cycle with fluorescent chemicals in DNA amplification reactions. Hybridization probes are a better choice than intercalating dyes such as SYBR Green in real-time quantitative PCR reactions. Fluorescently labeled probes can improve the efficiency, sensitivity, and specificity of real-time quantitative PCR results. Moreover, quantitative PCR can simultaneously detect multiple target genes in one reaction. During PCR amplification, a specific fluorescent probe is added while adding a pair of primers. The probe is a linear oligonucleotide, and the two ends are respectively labele...

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Problems solved by technology

However, due to the small size of echinococcosis, there is no obvious difference in the morphology of conventional microscopic examination, and the modification of the method is time-consuming and labor-intensive with low sensitivity. At present, there is no relevant molecular biological detection method

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