Level identification and bacterial strain typing integrated method of staphylococcus strain on the basis of single nucleotide polymorphism

Abstract

The invention discloses a level identification and bacterial strain typing integrated method of staphylococcus strain on the basis of single nucleotide polymorphism, as well as a target gene and a primer composition used for identification and typing. The method particularly includes the following steps: DNA extraction from to-be-detected bacterial strains, target gene PCR amplification, nucleic acid sequencing, sequence comparison, and clustering analysis, etc. On the basis of the method, a staphylococcus common target gene sequence databank is established, and the target gene sequence of the to-be-detected bacterial strain is compared and cluster-analyzed with a standard sequence in the databank, thus performing strain level identification and bacterial strain typing to the staphylococcus. The integrated method has high universality, is high in identification accuracy and has strong distinguishing capability, is simple in operations, is quick and effective, supplies a novel technology platform to identification and typing integrated detection of the staphylococcus, can be used in medicine quality supervision units and pharmaceutical enterprises, can be used for strain level identification and strain typing tracking of the staphylococcus, and has wide market prospect and huge economic and social benefits.

Description

technical field [0001] The invention relates to the technical field of drug safety microorganism detection, in particular to a primer composition for identification of staphylococcus species level and strain typing, and an integrated method for identification of staphylococcus species level and strain typing. Background technique [0002] With the improvement of drug production quality standards and national drug quality standards, the requirements for microbial inspection and identification levels in drug production quality control are getting higher and higher. The newly implemented "Chinese Pharmacopoeia" 2015 edition stipulates that the level of microbial identification depends on the situation, including species, genus identification and strain typing; In the process control, the laboratory should have the ability to find the source of pollution through the traceability analysis of the contaminated bacteria. [0003] Staphylococcus is widely distributed in nature. Trad...

AI Technical Summary

Problems solved by technology

At present, it has been targeted at Staphylococcus aureus [Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus [J]. J Clin Microbiol 38 (3): 1008-1015], Staphylococcus epidermidis [Improved multilocus sequence typing scheme for Staphylococcus epidermidis[J].J Clin Microbiol 45(2):616-619], Human Staphylococcus [Multilocus sequence typing and further genetic characterization of the enigmatic pathogen,Staphylococcushominis[J].PLoS One 8(6) :e66496] etc. have established corresponding MLST methods, but the target genes and primers used in these staphylococcal MLST typing methods are species-specific and cannot be used for typing of other staphylococcus strains

In addition, the application of the MLST method needs to be based on the identification of bacterial strains, which greatly reduces work efficiency and increases experimental costs.

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