Kit and method for detecting streptococcus suis type 2

A technology of kits and reagents, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems that are difficult to achieve high sensitivity and high specificity, and achieve high specificity, low detection cost, and high sensitivity Effect

Inactive Publication Date: 2012-07-25
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the defect that it is difficult to achieve high sensitivity and high specificity in the detection of Streptococcus suis type 2 at present, the invention provides a test kit and a detection method for Streptococcus suis type 2

Method used

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  • Kit and method for detecting streptococcus suis type 2
  • Kit and method for detecting streptococcus suis type 2
  • Kit and method for detecting streptococcus suis type 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The upstream primer A1 with the sequence 5'-CAAACGCAAGGAATTACGGTATC-3' (see Sequence Listing 2) and the 5' end modified with biotin and the sequence 5'-GAGTATCTAAAGAATGCCTATTG-3' (see Sequence Listing 3) were custom-purchased from Dalian Bao Biological Company. The downstream primer B1 and the probe C1 whose sequence is 5'-AAGTAGCAAGTAACCCTCCCGACA-3' (see Sequence Table 4) and whose 3' end is modified by digoxigenin.

[0056] Weigh 10g of beef extract, 20g of tryptone, 2g of glucose, 2g of sodium bicarbonate, 2g of sodium chloride, and 0.4g of disodium hydrogen phosphate, dissolve in 1000ml of distilled water, autoclave at 121°C for 15 minutes, and cool the culture medium to 45 At -50°C, add 10 mg of polymyxin E and 15 mg of nalidixic acid, and mix well to obtain a THB medium.

[0057] 0.5ml bacterium concentration is 10 / ml THB culture medium containing Streptococcus suis type 2 reference strain (purchased from ATCC company, product number is 43765) as sample, placed in...

Embodiment 2-8

[0065] Detect according to the same method as in Example 1, the difference is that according to Table 1, the corresponding culture medium containing the thalli of non-suis Streptococcus type 2 with a bacterial concentration of 10 / ml replaces the pig-containing pig with a bacterial concentration of 10 / ml. THB medium for Streptococcus type 2.

[0066] The optical density values ​​of the products after color development were measured by an enzyme-linked immunosorbent assay instrument are listed in Table 1.

[0067] Table 1

[0068] bacteria

[0069] *Nutritional Broth Medium was purchased from Guangdong Huankai Biotechnology Co., Ltd. (Product No. 022011)

Embodiment 9

[0075] The upstream primer A2 with the sequence of 5'-CGTATACTAATCTAGAGATTC-3' (see Sequence Table 5), the sequence of 5'-CTTATAAAGTTTGCAAC-3' (see Sequence Table 6) and the 5' end modified by biotin were purchased from Dalian Bao Biological Company. The downstream primer B2 and the probe C2 whose sequence is 5'-GTTTAAAAGAGAATGATAG-3' (see Sequence Table 7) and whose 5' end is modified by digoxigenin.

[0076] Detection was performed according to the same method as in Example 1, except that upstream primer A2, downstream primer B2 and probe C2 were used instead of upstream primer A1, downstream primer B1 and probe C1.

[0077] The optical density value (OD value) at 492 nm of the color-developed product measured by an enzyme-linked immunosorbent detector was 0.232.

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Abstract

The invention provides a kit and a method for detecting streptococcus suis type 2. The kit comprises an upstream primer, a downstream primer and a probe; the 5' end of the upstream primer and / or 5' end of the downstream primer are / is modified by a biotin; the 5' end and / or the 3' end of the of the probe are / is modified by digoxin; the upstream primer and the downstream primer can amplify the DNA molecule D or the fragment of the DNA molecule D through polymerase chain reaction to obtain a double-stranded DNA amplification product; the base sequence of the DNA molecule D is shown as in the sequence table 1; and the probe can hybridize flexibly with a biotin modified single-stranded DNA in the double-stranded DNA amplification product. The kit and the method have the advantages of high sensitivity, high specificity and low detection cost.

Description

technical field [0001] The invention relates to a kit and a method for detecting Streptococcus suis type 2. Background technique [0002] Streptococcus suis type 2, also known as Streptococcus suis type 2 (SS2), belongs to the R group of the Gram classification method, and is usually characterized by causing arthritis, meningitis, sepsis and bronchitis in weaned piglets , and can cause zoonotic diseases. Streptococcus suis type 2 is usually widely present in the respiratory tract of normal pigs, but the infection rate in tonsils of healthy pigs can also be as high as 76%. In recent years, Streptococcus suis type 2 has broken out many times in my country, not only causing huge losses in the breeding industry, but also infecting humans, causing great harm. [0003] At present, PCR method has been widely used to detect Streptococcus suis type 2, but its sensitivity and specificity need to be improved. [0004] For example, CN 1896279A discloses a method for detecting Strepto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 方勤美林天龙朱继昌
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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