LAMP primer composition used for detecting pathogenic microorganism of urogenital canal and related application thereof

A technology of urogenital tract and pathogenic microorganisms, applied in the field of LAMP primer composition, can solve the problems of difficult popularization, time-consuming, large subjective impact, etc., to improve detection sensitivity and specificity, make up for time-consuming and labor-intensive, and reduce labor intensity. Effect

Inactive Publication Date: 2017-08-29
SHANGHAI IGENETEC DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(3) Antigen detection: the sensitivity is not high, the specificity is poor, and it is greatly affected by the judgment level of the experimenter
However, this method is cumbersome to operate, time-consuming, expensive in equipment, highly technical, and not easy to carry out serological typing, so it is not easy to popularize in clinic
(3) The sensitivity of antigen detection test to diagnose Chlamydia trachomatis is only 70-90%, and the specificity is 83-99%.
(4) Serological tests are of litt

Method used

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  • LAMP primer composition used for detecting pathogenic microorganism of urogenital canal and related application thereof
  • LAMP primer composition used for detecting pathogenic microorganism of urogenital canal and related application thereof
  • LAMP primer composition used for detecting pathogenic microorganism of urogenital canal and related application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Design and preparation of the LAMP primer composition for detecting pathogenic microorganisms in the urogenital tract

[0032] Ribosomal DNA sequences exist in all organisms, have the same origin and function during evolution, and can reflect comparable evolutionary histories among species. The 16S rRNA gene is the DNA sequence corresponding to the coding rRNA on bacteria and exists in the genome of all bacteria. 16S rRNA is highly conserved and specific and the gene sequence is long enough. With the emergence of PCR technology and the continuous improvement of nucleic acid research technology, 16S rRNA gene detection technology has become a powerful tool for the detection and identification of pathogenic bacteria. The internal transcribed spacer (Internal Transcribed Spacer, ITS) of rDNA is a region between the 18S and 28S rDNA gene copy regions in eukaryotic rDNA, and the 5.8S rDNA is located between ITS1 and ITS2. The nucleotide sequence of 5.8S rDNA is h...

Embodiment 2

[0048] Example 2 Preparation and use of a kit for detecting genitourinary tract pathogenic microorganisms

[0049] Chlamydia trachomatis, Neisseria gonorrhoeae, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium, herpes simplex virus type 2, Candida albicans, Trichomonas vaginalis, human cytomegalovirus, syphilis, Proteus vulgaris, Clinical samples of Lactobacillus, Gardnerella vaginalis, Enterococcus and swabs of genital secretions of gonorrhea patients were obtained from Shanghai Dongfang Hospital.

[0050] 2.1 Preparation of kits for detecting pathogenic microorganisms in the genitourinary tract

[0051] The composition of the test kit for detecting urogenital tract disease-treating microorganism provided by the present invention is as follows:

[0052] 2.1.1 Isothermal amplification buffer

[0053] The solvent of the constant temperature amplification buffer is water, and the solute and concentration are as follows: 12mM dNTPs, 10×Isothermal Amplification ...

Embodiment 3

[0075] Example 3 Detection of human genital tract secretion swab samples using constant temperature amplification microfluidic chip

[0076] 3.1 Using the method of 2.1.3 in Example 2, the primer set for detecting Chlamydia trachomatis in Example 1, the primer set for detecting Neisseria gonorrhoeae, the primer set for detecting Ureaplasma urealyticum, and the primer set for detecting Primer set for Mycoplasma hominis, Primer set for detection of Mycoplasma genitalium, Primer set for detection of Herpes simplex virus type 2, Primer set for detection of Candida albicans, Primer set for detection of Trichomonas vaginalis, Surveillance Each primer in the primer set of the internal reference of the whole reaction is respectively fixed in reaction pool 1 and 17, reaction pool 2 and 18, reaction pool 3 and 19, reaction pool 4 and 20, reaction pool 9 and 25. Reaction pools 10 and 26, reaction pools 11 and 27, reaction pools 12 and 28, reaction pools 5, 13, 21 and 29; reaction pools 6...

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Abstract

The invention discloses an LAMP (loop-mediated isothermal amplification) primer composition used for detecting a pathogenic microorganism of the urogenital canal and a related application of the LAMP primer composition. The high-sensitivity high-accuracy LAMP primer composition used for detecting the pathogenic microorganism of the urogenital canal is provided, further, the LAMP micro-fluidic chip technique is applied to detecting the pathogenic microorganism of the sexual transmission urogenital canal, the characteristics of high throughput, rapidness, high sensitivity, and good specificity of a micro-fluidic chip are utilized to improve the detection speed, throughput and sensitivity in the detection of the pathogenic microorganism of the sexual transmission urogenital canal, so that the cost is greatly lowered, and the detection time is shortened.

Description

technical field [0001] The invention relates to the technical field of pathogen detection and diagnosis, in particular to a LAMP primer composition for detecting pathogenic microorganisms in the urogenital tract and related applications thereof. Background technique [0002] Sexually transmitted infection (Sexually Transmitted Infections, STI) is one of the most common public health problems in the world. The World Health Organization currently estimates that there are 250 million new cases of STIs worldwide each year. In my country, STIs mainly manifested as sexually transmitted urogenital infections (STUI) are increasing year by year. With the global spread of AIDS, HIV infection is showing a rapid growth trend in China. It is worth noting that STI can promote the heterosexual transmission of HIV / AIDS. The prevention and treatment of STUI is an important link in the prevention and control of HIV / AIDS, which has attracted people's attention. Especially nongonococcal ure...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04C12N15/11C12R1/01C12R1/35C12R1/36C12R1/725C12R1/90C12R1/93
CPCC12Q1/6844C12Q1/689C12Q1/6893C12Q1/6895C12Q1/705C12Q2600/166C12Q2531/119C12Q2565/629
Inventor 方雪恩王丽娟成引来黄皓孔继烈
Owner SHANGHAI IGENETEC DIAGNOSTICS CO LTD
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