Nucleotide sequence for MLVA (multiple-locus variable number tandem repeat analysis) typing detection of mycoplasma pneumoniae

A nucleotide sequence, Mycoplasma pneumoniae technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of different genotypes of the same standard strain, detection of different Mp genotypes, and different Mp genotypes And other issues

Inactive Publication Date: 2013-10-09
首都儿科研究所
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Problems solved by technology

Because Mpn1 is in a high-variation area, problems such as different genotypes of the same standard strain in different subculture batches, different Mp genotypes detected in the left and right lungs of the same patient, and different Mp genotypes before and after discharge from the same infected patient occurred during the detection process. Due to the different reaction conditions of each primer sequence, the detection process cannot be completed in the same reaction system

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  • Nucleotide sequence for MLVA (multiple-locus variable number tandem repeat analysis) typing detection of mycoplasma pneumoniae
  • Nucleotide sequence for MLVA (multiple-locus variable number tandem repeat analysis) typing detection of mycoplasma pneumoniae
  • Nucleotide sequence for MLVA (multiple-locus variable number tandem repeat analysis) typing detection of mycoplasma pneumoniae

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Embodiment Construction

[0035] The technical solutions of the present invention will be further explained below through the embodiments in conjunction with the accompanying drawings.

[0036] 1. Design of nucleotide primer sequence:

[0037] The sequence characteristics of the 8 primers are shown in Table 1.

[0038] Table 1. Primer Characteristics for MLVA-CE Genotyping

[0039]

[0040] 2. The application of the above primers to the MLVA genotyping of Mycoplasma pneumoniae, specific steps:

[0041] (1) Sample DNA extraction: The sample is from a patient suspected of mycoplasma infection, and the sample can be sputum, nasopharyngeal swab, alveolar lavage fluid, pleural effusion, cerebrospinal fluid, joint fluid, blood and other body fluids that can come from the patient; or clinical To isolate the cultured product, take 400 ul of the sample stock solution, centrifuge at 15,000 rpm for 15 minutes, and discard the supernatant. Add 50ul of Lysis Solution A (containing 5‰NP-40), and 2ul of Lysis S...

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Abstract

The invention relates to a nucleotide sequence for MLVA (multiple-locus variable number tandem repeat analysis) typing detection of mycoplasma pneumoniae, belonging to the field of biomedicines. According to the nucleotide sequence disclosed by the invention, a specific nucleotide primer sequence is designed against 4 VNTR (variable number of tandem repeats) regions, namely MLVA (multiple-locus variable number) 1, MLVA 2, MLVA 3 and MLVA4 and is used for MLVA detection of the mycoplasma pneumoniae, and genotyping is performed on the mycoplasma pneumoniae according to the combination of numbers of repeated sequences of the 4 VNTR regions. A large number of clinical specimens prove that the method disclosed by the invention can be directly used for performing stable genotyping on the specimens infected with the mycoplasma pneumoniae and used for development of Mp typing identification kits.

Description

technical field [0001] The invention relates to a nucleotide sequence for MLVA typing detection of Mycoplasma pneumoniae. 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 and bronchitis to severe interstitial pneumonia (primary atypical Pneumonia) and nervous system, cardiovascular system complications, etc. 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 great economic losses and social adverse effects on famili...

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
Inventor 孙红妹闫超薛冠华李少丽赵汉青冯燕玲
Owner 首都儿科研究所
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