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Mycoplasma pneumonia P1-RFLP gene typing and detecting primer and method

A technology of P1-RFLP and Mycoplasma pneumoniae, which is applied in the fields of medicine and biology, and can solve the problems of difficult amplification, laborious, large-scale detection, etc.

Inactive Publication Date: 2013-04-10
首都儿科研究所
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Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a kind of Mycoplasma pneumoniae P1-RFLP typing detection primer and method, respectively design 4 inner sets of primers to RepMP2 / 3 and RepMP4 sequences, by amplifying shorter fragments, can directly detect Mycoplasma pneumoniae Infected specimens are genotyped to solve the current problem that the P1-RFLP genotyping amplification fragment of Mycoplasma pneumoniae is relatively long, difficult to be amplified, time-consuming, laborious, and cannot be detected on a large scale in clinical practice

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  • Mycoplasma pneumonia P1-RFLP gene typing and detecting primer and method

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[0023] The technical solutions of the present invention will be further explained below through the embodiments in conjunction with the accompanying drawings.

[0024] According to the known RepMP4 and RepMP2 / 3 sequences of the P1 genes of the two international standard strains of Mp, analyze the region where the HaeIII restriction site is located, and design the inner set of primers for the region containing the restriction site in the middle of the sequence to both ends respectively. Specific amplification, the 2201bp of the full-sequence RepMP4 sequence and the 2501bp of the RepMP2 / 3 sequence, which were originally difficult to amplify, were divided into two shorter fragments for amplification respectively, and the amplification products of the RepMP4 sequence were 1150bp and 1061bp respectively; RepMP2 The amplified products of / 3 sequence were 1265bp and 1249bp respectively. Then these amplified products were digested with HaeIII endonuclease, and different types were jud...

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Abstract

The invention discloses a mycoplasma pneumonia P1-RFLP gene typing and detecting primer and method. The method comprises the following steps of: analyzing the region where a HaeIII enzyme digestion site is arranged according to the RepMP4 and RepMP2 / 3 sequences of two known Mp natural standard strain P1 genes; designing adjacent inner nested primers in middle parts of the sequences and regions containing the enzyme digestion site, and respectively carrying out specific amplification towards two ends; then, respectively mixing the inner nested amplification products, and then carrying out enzyme digestion by using the HaeIII incision enzyme; and judging different types according to different enzyme-digested product atlases. The problems that the typed and amplified fragment of a mycoplasma pneumonia P1-RFLP gene is longer and not easy to amplify, time and effort are wasted, and large-scale clinical detection can not be carried out are solved.

Description

technical field [0001] The invention belongs to the fields of biology and medicine, and in particular relates to a primer and a method for detecting mycoplasma pneumoniae P1-RFLP typing. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, Mp) is one of the important pathogens of community-acquired respiratory tract infection, and its clinical manifestations after infection can range from mild pharyngitis, bronchitis to severe interstitial pneumonia (primary atypical Pneumonia) and nervous system, cardiovascular system complications. The prevalence of Mp is cyclical, and the incidence peaks in different regions every 3-7 years. 10-30% of children's pneumonia cases are caused by Mp, and Mp pneumonia can be as high as 30-50% in the epidemic peak year, and the epidemic can last for 1-2 years. Since the beginning of the 21st century, there have been two global outbreaks of Mp infection in 2005-2007 and 2010-2012. The outbreak of Mp infection will have g...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 孙红妹薛冠华李少丽赵汉青冯燕玲
Owner 首都儿科研究所
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