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
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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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