Genital tract pathogen nucleic acid detection kit

A technology for detecting kits and pathogens, which is applied to the determination/inspection of microorganisms, microorganisms, and methods based on microorganisms. It can solve the problems of expensive, time-consuming, and false positives for supporting instruments, and achieve lower detection costs, less inconvenience for patients, and lighten the The effect of financial burden

Inactive Publication Date: 2019-03-12
MERLIN BIOMEDICAL (XIAMEN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The culture technique is tedious and time-consuming; the immunization technique requires specific antiserum
The superiority of PCR technology itself is beyond reproach, but due to many experimental steps, and PCR aerosol is very prone to contamination, resulting in serious false positive results, supporting equipment is expensive, consumables adopt an open design, and insufficient consideration is given to biological safety

Method used

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  • Genital tract pathogen nucleic acid detection kit
  • Genital tract pathogen nucleic acid detection kit
  • Genital tract pathogen nucleic acid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A nucleic acid detection kit for reproductive tract pathogens, comprising a first detection component, a second detection component, and a third detection component in a freeze-dried state that are simultaneously subjected to fluorescence quantitative PCR detection after adding a detection sample, and the first detection component is used to detect trachoma For Chlamydia and Neisseria gonorrhoeae, the second detection component is used to detect Ureaplasma urealyticum and Mycoplasma hominis, and the third detection component is used to detect Mycoplasma genitalium;

[0034] Among them, the first to third detection components are all made of trehalose, mannitol, PEG and detection reagent composition mixed and then freeze-dried, and the detection reagent composition of the first to third detection components are all made of 10×PCR buffer , dNTPs, detection primer pair, internal reference primer pair, detection probe, internal reference probe, MgCl 2 , BSA, r-Taq enzyme, U...

Embodiment 2

[0053] Detect respectively Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), Ureaplasma urealyticum (UU), Neisseria gonorrhoeae (NG) and Mycoplasma hominis (MH) pathogen (dry powder of pathogen) with embodiment 1 reproductive tract pathogen nucleic acid detection kit All were purchased from ATCC and reconstituted into 10 6 bacteria solution per mL), specifically, add 100-200 μL of bacteria solution to the first to third detection components, and place it in a fluorescent quantitative PCR instrument for amplification detection according to the following amplification procedure: pre-denaturation at 95°C for 3 minutes; Add 40 cycles, 95°C for 15s, 60°C for 30s (collect fluorescence)

[0054] The fluorescent PCR diagram for detecting Chlamydia trachomatis (CT) alone is as follows figure 1 and figure 2 Shown, the fluorescent PCR picture of detecting Mycoplasma genitalium (MG) alone image 3 and Figure 4 Shown, the fluorescent PCR picture of detecting Ureaplasma urealyti...

Embodiment 3

[0056] The nucleic acid detection kit for reproductive tract pathogens in Example 1 was used to detect the swab sample 1 and swab sample 2 of the patient's vaginal and cervical secretions from the Xiamen Maternal and Child Health Hospital, respectively, and soak the swab sample 1 and swab sample 2 with 1mL physiological saline , shake for 120s, and obtain sample stock solution 1 and sample stock solution 2.

[0057] Add 200-500 μL of sample stock solution 1 to the first to third detection components, cover the lid, put it into the matching automatic PCR analysis system, and run it for 40-60 minutes to obtain the following detection results, that is, Neisseria gonorrhoeae (NG) positive (Such as Figure 11 shown).

[0058] Add 200-500 μL sample stock solution 2 to the first to third detection components respectively, cover the lid, put it into the matching automatic PCR analysis system, and run it for 40-60 minutes to obtain the detection results as follows, that is, Mycoplasma...

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Abstract

The invention discloses a genital tract pathogen nucleic acid detection kit which comprises a first detection component, a second detection component and a third detection component which are freeze-dried and simultaneously perform fluorescent quantitative PCR (polymerase chain reaction) detection after detection samples are added. The first detection component is used for detecting Chlamydia trachomatis and Neisseria gonorrhoeae, the second detection component is used for detecting mycoplasma urealytium and mycoplasma hominis, and the third detection component is used for detecting mycoplasmagenitalium. The kit can simultaneously detect five nucleic acids such as the Chlamydia trachomatis, the Neisseria gonorrhoeae, the mycoplasma urealytium, the mycoplasma hominis and the mycoplasma genitalium in a genital tract, detection of five pathogenic microorganisms is integrated into a consumable, reagent stability is improved by the aid of a freeze-drying process, the kit is disposable whenbeing used by each user, only single sampling and one-step operation are needed, laboratory operators are greatly free, detection cost is reduced, inconvenience of a patient is decreased, and the financial burden of the patient is relieved.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a nucleic acid detection kit for reproductive tract pathogens. Background technique [0002] Infertility refers to the failure to conceive without taking contraceptive measures for one year and having normal sex life. Research shows that infertility afflicts approximately 16% of couples of reproductive age worldwide. Investigations at home and abroad have shown that the fallopian tube factor among biological factors is the primary factor causing infertility in China. The infection of reproductive tract pathogens is the main factor leading to infertility. Mycoplasma, Chlamydia and Neisseria gonorrhoeae are the main pathogenic bacteria of reproductive tract infection, which can cause urethritis in men, vaginitis in women, cervicitis, pelvic inflammatory disease and infertility. [0003] Sexually transmitted diseases caused by Chlamydia trachomatis (CT) ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/04C12R1/35C12R1/01
CPCC12Q1/689C12Q2600/16C12Q2600/166
Inventor 郑丽金朱伟灏沈义峰
Owner MERLIN BIOMEDICAL (XIAMEN) CO LTD
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