Method for detecting serotyping of streptococcus pneumoniae

Abstract

The invention provides a method for carrying out serotyping on streptococcus pneumoniae by using MLPA (multiplex ligation dependent probe amplification). The method comprises the following steps of: firstly, extracting a sample genome DNA of streptococcus pneumoniae to be detected; carrying out pre-amplification on the extracted genome DNA by using a PCR pre-amplification primer; carrying out hybridization on pre-amplification products of the genome DNA by using an MLPA probe; adding a corresponding fluorescence detecting probe in a reaction system; connecting hybridized MLPA probes by using ligase, and carrying out PCR amplification on the hybridized and connected MLPA probes by using an MLPA universal amplification primer; and analyzing PCR products by using a multicolor fluorescence dissolution curve, so that the serotyping on streptococcus pneumoniae can be realized. The detecting method disclosed by the invention is good in specificity and high in sensitivity; according to the method, 10 kinds of serotypes can be subjected to genotyping simultaneously just in 2-4 hours, so that multiple uncapping operations in traditional MLPA detection are avoided, and the pollution possibility is reduced, therefore, the method can meet high-throughput sample detection, and is especially applicable to inspection departments.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a detection technology for serotype typing of Streptococcus pneumoniae, and specifically relates to a detection method for serotype typing of Streptococcus pneumoniae, which is suitable for qualitative detection of the serotype of Streptococcus pneumoniae. Background technique [0002] Streptococcus pneumoniae (Streptococcus pneumonia), also known as Diplococcus pneumoniae or pneumococcus, is positive for Gram staining and generally arranged in a double spherical shape under a microscope; it has a capsule structure, and the capsule is the main virulence factor of Streptococcus pneumoniae. According to the structure and composition of capsular polysaccharides, Streptococcus pneumoniae can be divided into 46 serogroups and more than 90 different serotypes. About 20 serotypes / groups are associated with common pneumococcal invasive disease, and most serotypes are harmless to humans. At pr...

AI Technical Summary

Problems solved by technology

However, the traditional MLPA technology needs to be opened many times during the operation, which increases the risk of nucleic acid aerosol contamination, and the result detection requires an expensive capillary electrophoresis sequencer, which hinders its application in clinical testing

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