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Treponema pallidum (TP) nucleic acid testing kit

A technology of Treponema pallidum and detection kit, which is applied in the direction of microorganisms, biochemical equipment and methods, and methods based on microorganisms, can solve the problems that cannot be used to observe the curative effect, and achieve good product stability, high sensitivity, and detection results. reliable and accurate results

Inactive Publication Date: 2012-12-12
泰普生物科学(中国)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method detects anti-Treponemal pallidum IgG or IgM antibodies in the serum. Even if the patients undergo adequate treatment, the antibodies can still exist for a long time, or even persist for life, and the seroreaction continues to be positive. Therefore, it cannot be used to observe the curative effect

Method used

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  • Treponema pallidum (TP) nucleic acid testing kit
  • Treponema pallidum (TP) nucleic acid testing kit
  • Treponema pallidum (TP) nucleic acid testing kit

Examples

Experimental program
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Effect test

preparation example Construction

[0064] The preparation method of the extract A is: take 684-1368g sucrose, 7.874-10.297g tris (Tris), 250-350mL polyethylene glycol octyl phenyl ether (TritonX-100), mix well , adjust the pH value to 7.5-8.0 with hydrochloric acid, dilute the volume to 3000mL with pure water, and take 3mL per person;

[0065] The preparation method of the extract B is: take 0.05-0.15g polystyrene matrix ion exchange resin (Chelex100), 0.25-0.75mL TritonX-100, 0.04-0.08g Tris, hydrochloric acid to adjust the pH value to 7.8-8.3, pure Dilute water to 50mL, take 0.05mL per person.

[0066] The kit also includes positive and negative quality controls for Treponema pallidum nucleic acid detection.

[0067] The preparation method of the positive quality control product is as follows: a) collect the sample that is detected as TP positive by sequencing, b) obtain the plasmid through PCR amplification and T vector cloning, c) transform the plasmid into Escherichia coli, d) strain culture, e ) Collect...

Embodiment 1

[0072] Table 1: Kit Composition

[0073]

[0074] The preparation method of extract A is as follows: Take 684g sucrose, 7.874g Tris (Tris), 250mL polyethylene glycol octylphenyl ether (TritonX-100), mix well, and adjust the pH value to 7.5- 8.0, dilute pure water to 3000mL, take 3mL per person;

[0075] The preparation method of extract B is: take 0.05g polystyrene matrix ion exchange resin (Chelex100), 0.25mL TritonX-100, 0.04g Tris, hydrochloric acid to adjust the pH value to 7.8-8.3, pure water to 50mL, take each Serving 0.05mL.

[0076]

[0077] Table 2: TP-PCR Reaction Solution Recipe

[0078] serial number components initial concentration quantity 1 pure water - 25 μL 2 PCR buffer 10x 5μL 3 MgCl 2 25mM 7μL 4 dNTP - 1μL 5 L-pri-T1 10μmol 1μL 6 L-pri-T2 10μmol 1μL 7 L-pri-I1 10μmol 0.5μL 8 L-pri-I2 10μmol 0.5μL 9 L-pro-T1 10μmol 1μL 10 L-pro-I1 10μmol 0.5...

Embodiment 2

[0152] Embodiment 2 The use of kit of the present invention

[0153] 1. Reagent preparation

[0154] 1. Prepare materials

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Abstract

A TP nucleic acid testing kit mainly comprises a polymerase chain reaction (PCR) reaction reagent and a DNA extraction reagent. The testing kit is characterized in that the PCR reaction reagent comprises an upstream outer primer of specifically amplified TP nucleic acid, a downstream outer primer of the specifically amplified TP nucleic acid, a probe for detecting the TP nucleic acid, wherein the base sequence of the upstream outer primer is 5'-GGTCGATGTGCAAATGAGTGTT-3; The base sequence of the downstream outer primer is 5'-CCGACTTCAATACGGAGTTCAC-3'; the base sequence of the probe is 5'-ACTAGCCCTCCCTTCTACCTGAGATAAG-3'; and the 5'end of the probe is marked through fluorescent dye, and the 3'end of the probe is marked through quenching fluorescent dye. The testing kit has the advantages of being high in sensitivity, specificity and throughput, capable of detecting the TP rapidly and accurately, and suitable for clinical application and the like.

Description

technical field [0001] The invention relates to a disease pathogen gene detection technology, in particular to a treponema pallidum nucleic acid detection kit. Background technique [0002] Treponema pallidum (Treponema pallidum, TP) belongs to Treponema pallidum subsp. pallidum, which is the pathogen that causes human syphilis. Once infected, the spirochete spreads quickly throughout the body and can invade almost every organ of the human body. It manifests a variety of clinical manifestations similar to many diseases. There are three ways of infection of syphilis: direct contact, indirect contact and placental infection, among which sexual direct contact is the main way of syphilis infection. In addition, congenital syphilis caused by placental infection is increasingly attracting national attention. [0003] The earliest discovery and discussion of syphilis patients was in North America. It was introduced to Guangdong, my country via India in 1505, and it has been more...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/01
Inventor 李旭新杨玉芬
Owner 泰普生物科学(中国)有限公司
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