Kit for simultaneously detecting neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid, and application of kit

A technology for Neisseria gonorrhoeae and Chlamydia trachomatis, which is applied in the field of biological detection and can solve problems such as easy pollution

Active Publication Date: 2020-06-19
武汉中帜生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now commonly used STD pathogenic acid detection is based on fluorescent PCR method, these methods need special PCR amplification conditions, special laboratory and fluorescent quantitative PCR instrument, and it is easy to produce pollution during the detection process

Method used

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  • Kit for simultaneously detecting neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid, and application of kit
  • Kit for simultaneously detecting neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid, and application of kit
  • Kit for simultaneously detecting neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid, and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] [Example 1] Sensitivity Test

[0098] Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum nucleic acid detection kit can specifically detect Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum, the common serum of Neisseria gonorrhoeae and Chlamydia trachomatis was selected in this study Type F, common serotype 3 of Ureaplasma urealyticum, the minimum detection limit was determined. See the table below for information:

[0099] Table 1 NG pathogen information

[0100] Pathogen name Bacterial Strains (ATCC) Corresponding unit NG 19424 >10 6 cfu / mL

CT-F VR-346 1.8×10 6 IFU / mL

UU 27815 10 7 CCU / vial

[0101] Each gradient virus dilution was repeated 3 to 5 times, and each copy was repeated 20 times. The virus level with a positive detection rate of 90% to 95% was used as the minimum detection limit. The detection results were as follows:

[0102] NG minimum detection limit det...

Embodiment 2

[0122] [Example 2] Specificity Verification

[0123] 1. Test Strains

[0124] After extracting nucleic acids from different microorganisms, they are tested to verify the specificity of the design of the primers and probes of the kit of the present invention. Relevant pathogens and titers are as follows:

[0125] Table 5 Specificity verification test strain information

[0126] microorganism concentration Mycoplasma hominis 1.0×10 7 CCU / mL

Mycoplasma genitalium 1.0×10 7 CCU / mL

Herpes simplex virus type I 1.0×10 6 TCID50 / mL

Herpes simplex virus type II 1.0×10 6 TCID50 / mL

Cytomegalovirus 1.0×10 6 TCID50 / mL

papillomavirus type 18 1.0×10 6 copies / mL

Candida albicans 2.8×10 6 TCID50 / mL

Hepatitis A virus 3.2×10 9 TCID50 / mL

Hepatitis B virus 8.89×10 8 TCID50 / mL

Staphylococcus aureus (24~40)×10 8 cfu / mL

Escherichia coli (12~14)×10 8 cfu / mL

[0127] ...

Embodiment 3

[0134] [Example 3] Verification of samples

[0135] 1. Clinical sample information

[0136] The tests came from a total of 320 samples from Wuhan First Hospital, including 189 male samples and 131 female samples, accounting for 59.06% and 40.94% respectively. Among the 452 samples, the oldest patient was 67 years old, the youngest was 16 years old, the average age was 31.25 years old, the standard deviation was 33.5 years old, and the median was 29 years old. The diagnosis results of the enrolled patients were all related to reproductive tract infection.

[0137] 2. Test results

[0138] (1) Neisseria gonorrhoeae test results

[0139] During the detection, the test kit of the present invention and a certain commercialized detection Neisseria gonorrhoeae fluorescent quantitative PCR kit were used to detect the samples simultaneously, and the detection results were summarized into four tables, as follows:

[0140]

[0141] For the 5 inconsistent samples, the "gene sequenci...

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Abstract

The invention discloses a kit for simultaneously detecting neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid, and application of the kit. After a collected sample issubjected to pyrolysis through a cell lysis solution, pathogen nucleic acid is released, and through reverse transcription and transcription processes, the amplification of a pathogen nucleic acid fragment is realized. An amplified RNA (Ribonucleic Acid) product is added into a micropore coated with a coating probe, and meanwhile, a specific probe and an enlargement probe are added, wherein the coating probe can be bound with one end of the specific probe CES to fix the amplified product RNA. One end of a specific probe LES is bound to the RNA product, and the other end is bound with the enlargement probe to realize signal enlargement. The kit does not require RNA extraction, is unlikely to be polluted during detection, is high in sensitivity and specificity, and can be widely used for detecting the neisseria gonorrhoeae, chlamydia trachomatis and ureaplasma urealyticum nucleic acid.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for jointly detecting nucleic acids of Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum based on double amplification (RNA constant temperature amplification + multi-biotin signal amplification) technology and its application. Background technique [0002] Sexually transmitted diseases refer to a group of infectious diseases mainly transmitted by sexual contact, referred to as sexually transmitted diseases. Common venereal pathogens include viruses, bacteria, mycoplasma, and chlamydia. Urogenital tract infections are mainly caused by Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum, and the three are often co-infected. [0003] Sexually transmitted diseases are a high incidence worldwide, and the incidence rate is increasing year by year. Infection occurs mostly in people who are sexually active, have unclean sexu...

Claims

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Application Information

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IPC IPC(8): C12Q1/682C12Q1/6834C12Q1/689C12Q1/04
CPCC12Q1/682C12Q1/6834C12Q1/689C12Q2600/166C12Q2531/119C12Q2547/101C12Q2545/101C12Q2545/113C12Q2563/103C12Q2521/107
Inventor 李先强姜昕黄永伟王琳琳薛金辉
Owner 武汉中帜生物科技股份有限公司
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