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Detecting and parting method of chlamydia trachomatic PCR RLB and used primer, probe

A technology of Chlamydia trachomatis and typing method, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc. It can solve the problems of repeated steps, inability to detect and identify multiple types of CT mixed infection at the same time, and long time consumption

Inactive Publication Date: 2006-01-25
深圳市慢性病防治院
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Problems solved by technology

However, current studies believe that the nucleic acid sequence of the VD2 region of the CTompl gene is a conserved nucleic acid sequence with the most differences among different types
Although the PCR-RFLP diagnostic method based on the VD2 region of the ompl gene can accurately monitor each type of CT, it cannot detect and identify mixed infection of multiple types of CT at the same time, and it also has the disadvantages of repeated steps and long time consumption
DNA sequencing is also not suitable for identification of polytype CT co-infection

Method used

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  • Detecting and parting method of chlamydia trachomatic PCR RLB and used primer, probe
  • Detecting and parting method of chlamydia trachomatic PCR RLB and used primer, probe
  • Detecting and parting method of chlamydia trachomatic PCR RLB and used primer, probe

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Embodiment Construction

[0079] Attached below picture with Embodiment further elaborates the present invention:

[0080] 1. Primer and Probe Design

[0081] According to the general principle of PCR amplification oligonucleotide design and the principle that the Tm value of oligonucleotide melting temperature is similar and the size of corresponding amplified fragments can be distinguished in ordinary agarose gel electrophoresis, use WebANGIS to compare CT15 species Type omplVD2 sequence, design 3 pairs of CT primers, 3 kinds of specific probes and 15 types of specific probes. In order to improve the sensitivity of the detection method, we designed 3 pairs of universal primers (CP24b, CP27b; CTSb, CTSNb, CTANb, CTAb). The 5' of the primer is biotin-labeled, and the 5' of the probe is amino-labeled. The sequences of the primers are as follows:

[0082] Oligonucleotide 101 (CP24b): 5'-GGGATTCCTGTAACAACAAGTCAGG-3'

[0083] Oligonucleotide 102 (CP27b): 5'-CCTCTTCCCCAGAACAATAAGAACAC-3'

[0084] O...

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Abstract

The invention discloses a subtype method to testing Chlamydia trachomatis PCR-RLB, and the primer and probe. Three pair oligonucleotide primer, three strain specificity probes, 15 specific type specificity probes are designed aimed at the DNA sequence of kinds of Chlamydia trachomatis. It supplies a method to testing CT infection while CT diagnosing genitourinary tract, taking subtype accurately to CT, and diagnosing mixed infection. It lays a foundation for CT pathogenesis research.

Description

[technical field] [0001] The invention relates to a novel molecular biology method for detecting and typing the chlamydia trachomatis; in particular, it relates to a method for detecting the chlamydia trachomatis by PCR-RLB and its typing, as well as primers and probes used therein. [Background technique] [0002] Chlamydia trachomatis (Chlamydia Trachomatis, CT) is a common venereal pathogen, belonging to a kind of Chlamydia. Chlamydia is an intracellular parasite with a unique growth cycle that can only survive inside the host cell. Extracellular Chlamydia primary body EBs are taken up into cells to form vacuolar inclusion bodies, and then transform into reticulosome RBs. RB has the ability to reproduce, but not to infect; the RB that reproduces in the cell quickly transforms into EB, which lacks the ability to reproduce but has the ability to infect. It penetrates the inclusion body, destroys the cell wall, and reaches the outside of the cell. The World Health Organizat...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 熊礼宽程锦泉周华孔繁荣玲.格伯特
Owner 深圳市慢性病防治院
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