A primer set, probe set and kit for synchronous detection of pathogens associated with multiple reproductive tract infections

A technology for reproductive tract infection and primer probes, which is applied to the field of primer compositions for pathogens and their kits, can solve the problems of low sensitivity, low sensitivity, and large impact on results, and achieves strong primer specificity and high reverse transcription efficiency. , check the overall effect

Active Publication Date: 2021-03-23
爱科睿特生物医疗科技(南京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus culture detection method has the characteristics of high specificity and is the "gold standard" for clinical diagnosis, but it is time-consuming, low-sensitivity, and requires high experimental operation; bacterial culture can detect drug resistance, but the sample is easy to contaminate, which affects the results. The impact is relatively large; the immunological detection of pathogens in reproductive tract infections often uses enzyme-linked immunosorbent assay. Clinical reference value; Fluorescent quantitative PCR technology has the advantages of high sensitivity, rapid detection, and relatively low cost, and has been widely used in clinical diagnosis
[0007] However, the specificity of dye-based fluorescent quantitative PCR is relatively poor, and different target products cannot be quantified separately. Multiplex fluorescent quantitative PCR is a multiple joint detection developed on the basis of it, which makes up for these shortcomings and realizes simultaneous detection in the same reaction tube. Amplification and specific detection of multiple gene sequences

Method used

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  • A primer set, probe set and kit for synchronous detection of pathogens associated with multiple reproductive tract infections
  • A primer set, probe set and kit for synchronous detection of pathogens associated with multiple reproductive tract infections
  • A primer set, probe set and kit for synchronous detection of pathogens associated with multiple reproductive tract infections

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of a multiplex fluorescent quantitative PCR kit for the detection of 15 reproductive tract pathogens

[0046] The multiplex fluorescent quantitative PCR amplification kit used in the detection of 15 kinds of reproductive tract pathogens of the present invention, the composition of the PCR amplification kit is shown in Table 1.

[0047] Table 1 Composition list of PCR amplification kit

[0048]

[0049] The kit of the present invention can be used to detect 15 kinds of reproductive tract pathogens: herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), group B streptococci (SAg), atopobium (AV), common Revobacterium (PV), Megasphaerum (ME), Motonia (MB), Neisseria gonorrhoeae (NG), Gardnerella (GV), Trichomonas vaginalis (TV), Treponema pallidum (TP) , Candida albicans (CA), Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), Mycoplasma hominis (MH) nucleic acid.

[0050] The design of 15 kinds of reproductive tract patho...

Embodiment 2

[0075] Embodiment 2: the detection method of multiplex fluorescent quantitative PCR kit

[0076] 1. Detection kit

[0077] The multiplex fluorescent quantitative PCR kit prepared in Example 1 for the detection of 15 kinds of reproductive tract pathogens.

[0078] Detection object

[0079] 2.1 Preparation of internal standard granule normal saline containing housekeeping gene (hereinafter referred to as "internal standard normal saline"): Measure 800mL of normal saline (0.9% sodium chloride solution) into a 1000mL volumetric flask in the reference substance preparation room in the clean work area , adding 5.0×10 9 Copies / mL pathogen plasmid or particle 1mL, dilute to 1000mL with normal saline, turn over to mix well, so that the final concentration is 5.0×10 6 copies / mL. Transfer the above solution to a 1000mL beaker and divide it for use.

[0080] 2.2 7 samples to be tested were negative

[0081]Collect cervicovaginal secretion samples such as HPV16, HPV18, HPV51, ciliat...

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Abstract

The invention discloses a primer set, a probe set and a kit for synchronously detecting pathogens related to various reproductive tract infections. The sequences of the primer composition and the probe are SEQ ID NO: 1-45, each sequence of the probe set The two ends of each have a fluorescent group and a quencher group respectively. The invention discloses a kit, which includes the primer set and probe set, pathogen marker standard solution and PCR reaction commonly used Enzymes and reagents, the kit of the present invention can detect 15 pathogens related to reproductive tract infection at the same time, the kit of the present invention has high reverse transcription efficiency, strong primer specificity, high sensitivity, comprehensive, fast and accurate detection, and is widely used , the test results can be used for clinical diagnosis and treatment.

Description

technical field [0001] The invention relates to the technical field of nucleic acid amplification, in particular to a primer composition and a kit for synchronously detecting multiple pathogens related to reproductive tract infections. Background technique [0002] In recent years, the number of patients with genitourinary tract infections has been increasing. Epidemiological surveys of married women show that the prevalence of such infections has become the first among gynecological diseases. The pathogens that cause reproductive tract infections are various and complex, and there are certain asymptomatic infections, which cause many difficulties in the diagnosis and treatment of diseases. Genital tract pathogen infection usually leads to the following conditions: urinary system infection, vaginitis, pelvic inflammatory disease, etc. Reproductive tract pathogen infection will increase the incidence of male and female infertility, ectopic pregnancy, miscarriage, premature b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6895C12Q1/689C12Q1/6851C12Q1/14C12Q1/04
CPCC12Q1/6851C12Q1/689C12Q1/6895C12Q1/705C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/107C12Q2521/107C12Q2545/113
Inventor 闫泉熊永清王洪
Owner 爱科睿特生物医疗科技(南京)有限公司
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