Multiple PCR detection kit for virulence factors of streptococcus suis and detection method thereof

A Streptococcus suis virulence factor and a Streptococcus suis virulence factor technology, which is applied to the multiple PCR detection kit of Streptococcus suis virulence factor and its detection field, and can solve the problem of inability to detect types 7 and 9 of Streptococcus suis , pathogenicity and other issues, to achieve the effect of reducing the probability of false negative and missed detection, strong specificity, stability and reliability

Inactive Publication Date: 2010-08-25
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] After searching the literature of the prior art, it was found that the Chinese invention patent application with the publication number CN1896279A disclosed a triple PCR rapid detection kit for Streptococcus suis type 2, but the kit could not detect the presence of Streptococcus suis types 7 and 9. Typing
In recent years, although there have been no outbreaks of streptococcosis suis caused by SS7 and SS9, there have been reports of SS7 or SS9 isolated from sick pigs at home and abroad. Further studies have shown that SS7 and SS9 are pathogenic, and when mixed infection Play a synergistic role in SS2 pathogenicity or outbreaks, so the development can cover 7 major virulence factors of Streptococcus suis (mrp, epf, sly, gdh, gapdh, orf2 and fbps) and 3 major pathogenic serotypes of Streptococcus suis (type 2, 7, 9) multiplex PCR detection kit is very necessary

Method used

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  • Multiple PCR detection kit for virulence factors of streptococcus suis and detection method thereof
  • Multiple PCR detection kit for virulence factors of streptococcus suis and detection method thereof
  • Multiple PCR detection kit for virulence factors of streptococcus suis and detection method thereof

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Embodiment 1

[0060] The kits and detection methods involved in this embodiment are as follows:

[0061] 1. Kit

[0062] The multiplex PCR detection kit of this embodiment includes 10 cryopreservation tubes, 50 PCR reaction tubes and instructions. The 10 cryopreservation tubes are specifically: cryopreservation tubes code-named M (2), C2, C7, C9, D, H, P1, P2, and P3, and cryopreservation tube M is equipped with 2×PCR core reagent premix substances (dNTPs, Taq DNA polymerase mix and MgCl 2 solution, commercially available), cryopreservation tubes C2, C7 and C9 are equipped with 30 μl of Streptococcus suis type 2, 7, and 9 positive control DNA solutions respectively, and cryopreservation tube D is equipped with 0.5ml of DL2000 molecular weight reference solution (commercially available), Cryotube H filled with 1.5ml of ddH 2 O, cryopreservation tubes P1, P2, and P3 are respectively equipped with dry powder mixtures of primers used in PCR systems I, II, and III; all cryopreservation tubes ...

Embodiment 2

[0105] The materials involved in this embodiment are as follows:

[0106] Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC25922), Streptococcus agalactiae (ATCC10556), Enterococcus faecalis (ATCC29212), and Campylobacter jejuni (ATCC33560) can be purchased from commercial companies such as Shanghai Fumeng Gene Biotechnology Co., Ltd. Company, company address: Fudan University, No. 229, Handan Road, Shanghai.

[0107] The Sichuan isolate ZY05719 (Streptococcus suis type 2) and the Jiangsu isolate HA9801 (Streptococcus suis type 2) have been published in many articles, such as "Shen Yan, Hua Xiuguo, Cui Li, etc. Two strains of Streptococcus suis type 2 Effects of representative isolates on some immune indicators after infection of miniature pigs[J]. Journal of Animal Husbandry and Veterinary Medicine, 2008, 39(12); 1721-1730", relevant strains can be obtained through open commercial channels, such as Qingdao China Animal Center for Hygiene and Epidemiology, Address: N...

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Abstract

The invention discloses a multiple PCR detection kit for virulence factors of streptococcus suis and a detection method thereof, and belongs to the technical field of biology. The multiple PCR detection kit comprises nucleic acid shown by base sequences, such as SEQ ID NO:1 to SEQ ID NO:20. The detection method for the multiple PCR detection kit comprises the following steps of: 1, providing DNA of samples to be detected; 2, taking the kit, amplifying the DNA of the samples to be detected by adopting the conventional PCR method, detecting an amplified result by adopting an agarose gel electrophoresis method, and judging according to the result; and by using positive DNA of streptococcus suis 2 type as contrast, if the contrast proves that not all the strips are amplified, re-detecting, and if the contrast proves that all the strips are amplified, judging the result. The multiple PCR detection method established by the invention is specific and sensitive, and has the advantages of simpleness, convenience and quickness. The multiple PCR detection kit has reliable stability, and can be used for rapid detection of clinical samples and survey research on molecular epidemiology for the streptococcus suis.

Description

technical field [0001] The invention relates to a multiplex PCR detection kit and a detection method thereof in the field of biotechnology, in particular to a multiplex PCR detection kit and a detection method thereof for virulence factors of Streptococcus suis. Background technique [0002] Streptococcus suis (SS) is an important zoonotic pathogen that is widely distributed throughout the world, causing meningitis, arthritis, endocarditis, sepsis, pneumonia, and sudden death in weaned piglets. Can infect humans. According to the different antigenicity of capsular polysaccharide, it can be divided into 35 serotypes (types 1-34 and 1 / 2). SS2, SS7, SS9 are the most important pathogenic strains, especially SS2 is the most popular and the most pathogenic. SS2 was isolated and identified for the first time in Guangdong Province in 1991, followed by outbreaks of human infection with SS2 in parts of Jiangsu and Sichuan in 1998 and 2005, resulting in death. [0003] Streptococcus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/46
Inventor 崔立华修国郑升博朱爱玲杨志彪
Owner SHANGHAI JIAO TONG UNIV
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