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Method for detecting and identifying 11 sexual transmission relevant microorganisms simultaneously

A detection method and microbial technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of high detection cost, limited number of pathogens, and low throughput of detection samples to achieve sensitivity high effect

Active Publication Date: 2019-09-17
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many defects in these platforms, such as high detection cost, low throughput of detection samples, limited number of pathogens detected by a single reaction, etc.

Method used

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  • Method for detecting and identifying 11 sexual transmission relevant microorganisms simultaneously
  • Method for detecting and identifying 11 sexual transmission relevant microorganisms simultaneously
  • Method for detecting and identifying 11 sexual transmission relevant microorganisms simultaneously

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1, detection method

[0074] In the method for highly sensitive detection and / or identification of sexually transmitted infection microorganisms provided by the embodiments of the present invention, 11 common clinical sexually transmitted infection microorganisms to be detected include: Trichomonas vaginalis, herpes simplex virus type 1, herpes simplex virus type 2, Neisseria gonorrhoeae, Chlamydia trachomatis, Treponema pallidum, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum, and Haemophilus ducreyi. Implementation of the present invention comprises the steps:

[0075] (1) For each microorganism to be detected, a gene that is specific among species and conserved within a species is selected as the target gene for detection. Firstly, the gene sequences of representative strains of each microorganism that had been fully annotated as reference sequences were downloaded from the GenBank database. The selected microorganism...

Embodiment 2

[0092] Embodiment 2, test kit

[0093] The test kit provided by the invention is used to detect the application in 11 kinds of sexually transmitted related microorganisms, comprising the following steps:

[0094] 1) Nucleic acid extraction: Use a nucleic acid extraction kit to extract the secretion or urine sample from the test.

[0095] 2) Multiplex PCR reaction: Cooperate with a special amplification reaction system, after a round of multiplex PCR, the gene amplification of multiple entries is realized. Uracil N glycosylase (UNG enzyme) technology was introduced in the PCR process. In the first PCR reaction system, dUTP was used instead of dTTP in conventional PCR to incorporate a large amount of dU into the product. Before performing PCR amplification again, the residual contamination of PCR products can be eliminated by treating the PCR mixture with UNG enzyme. Since the UNG enzyme can be inactivated in one step of denaturation in the PCR cycle, it will not affect the ne...

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Abstract

The invention provides a method for detecting and identifying 11 sexual transmission relevant microorganisms simultaneously. According to the method disclosed by the invention, 11 sexual transmission relevant microorganisms are detected through a multiple PCR-mass spectrography and dual genes. The detection method comprises the following steps of (1) designing a primer; (2) performing a multiplex PCR amplification reaction; (3) performing shrimp alkaline phosphatase (SAP) treatment; (4) performing a basic group extension reaction; (5) performing resin desalination; and (6) performing mass spectrometric detection.

Description

technical field [0001] The invention relates to a method for detecting microorganisms, in particular to a method for detecting 11 kinds of microorganisms related to sexual transmission by multiplex PCR-mass spectrometry. Background technique [0002] Sexually transmitted infections (STIs) are one of the major causes of global public health and healthcare burden. STIs can lead to fetal / neonatal death, infertility, and genital tumors, and also greatly increase the risk of HIV infection. According to the report of the World Health Organization (WHO), there are approximately 357 million new STI cases worldwide each year, including chlamydia infection (131 million), gonorrhea (78 million), syphilis (05 million) and vaginal trichomoniasis (143 million). While many of these infections are preventable and treatable, mortality and morbidity associated with STIs remain high. Many factors contribute to the ongoing public health burden of STIs, the most important of which is delayed t...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6893C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/705C12Q1/689C12Q1/6893C12Q1/686C12Q2600/16C12Q2600/166C12Q2537/143C12Q2521/525C12Q2533/101C12Q2565/627C12Q2545/113Y02A50/30
Inventor 彭俊平郭军华修乐山
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI