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Haemophilus parasuis indirect hamagglutination detection reagent

A technology for Haemophilus suis and detection reagents, applied in the field of veterinary detection reagents, can solve the problems such as the lack of intellectual property rights of such products, the inability to really detect sensitized blood cells, and false negatives, so as to increase the reaction range and sensitivity, operation Simple, stable effect

Active Publication Date: 2015-03-18
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the IHA tests used by various institutions are only aimed at the detection of one or several serotypes of Haemophilus parasuis, and the antigen used to prepare sensitized blood cells is a single strain antigen of a certain serotype [Shi Bi et al., Chinese Veterinary Science, 2007,37(11); Liu Maojun et al., Porcine Science, 2007,12] or two serotype strain antigens [Miniats et al., Can J Vet Res, 1991,55(1); Tadjine et al., J Clin Microbiol, 2004, 42(2); Wei Zigong et al., Chinese Veterinary Science, 2006, 36(9)], so false negatives or the possibility of missed detection often occur in clinical testing
In addition, it has been verified by the inventor many times that the sensitized blood cells prepared with the antigens of certain two serotype strains cannot really detect all serotypes of Haemophilus parasuis serum, but can only detect the serum of this serotype, and its effective Individual serotype sera that cross-react
At present, there is no intellectual property rights for such products

Method used

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  • Haemophilus parasuis indirect hamagglutination detection reagent
  • Haemophilus parasuis indirect hamagglutination detection reagent
  • Haemophilus parasuis indirect hamagglutination detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The antigens used in the preparation of Haemophilus parasuis indirect hemagglutination detection reagents in this example are serotype 1 Haemophilus parasuis N4 strain, serum type 4 Haemophilus parasuis FS0307 strain, and serum type 5 Haemophilus parasuis Nagasaki strain and serum type 13 Haemophilus parasuis IA-84-17975 strain, the antigenic protein concentration of each bacterial strain is 20ug / mL, and the preparation process is as follows:

[0018] 1. Culture of Haemophilus parasuis

[0019] Take one tube each of lyophilized and preserved Haemophilus parasuis serum type 1 (strain N4), type 4 (strain FS0307), type 5 (strain Nagasaki) and type 13 (strain IA-84-17975), add 0.5 mL TSB basal medium, after mixing, take 0.1mL of bacterial liquid and spread evenly on TSA solid medium (containing 5% serum and 0.01% NAD), and incubate at 37°C for 24-48h. Then pick 3 to 5 single colonies and inoculate them in 5 mL of TSB liquid medium (containing 5% serum and 0.01% NAD), and c...

Embodiment 2

[0047] The antigens used in the preparation of Haemophilus parasuis indirect hemagglutination detection reagents in this example are serotype 1 Haemophilus parasuis N4 strain, serum type 4 Haemophilus parasuis FS0307 strain, and serum type 5 Haemophilus parasuis Nagasaki Strain and serum type 13 Haemophilus parasuis IA-84-17975 strain, the antigenic protein concentration of each strain is 40ug / mL;

[0048] Using the above-mentioned batches of Haemophilus parasuis hemagglutination detection reagents prepared in this example, according to the same detection procedure as in Example 1, the negative serum, 15 serotypes and 2 indeterminate positives prepared and preserved in the testing laboratory were tested. Serum samples, the test results are as follows:

[0049] Table 5 The results of negative and positive samples detected by indirect hemagglutination reagent for multiple serotypes of Haemophilus parasuis

[0050] Type 1

Type 2

type 3

Type 4

Type 5

...

Embodiment 3

[0061] In this example, the antigens used in the preparation of Haemophilus parasuis indirect hemagglutination detection reagents are serotype 1 Haemophilus parasuis strain N4, serum type 4 Haemophilus parasuis strain FS0307, ​​and serum type 5 Haemophilus parasuis XX0306 Strain and serum type 13 Haemophilus parasuis IA-84-17975 strain, the antigenic protein concentration of each strain is 20ug / mL;

[0062] Using the above-mentioned batches of Haemophilus parasuis hemagglutination detection reagents prepared in this example, according to the same detection procedure as in Example 1, the negative serum, 15 serotypes and 2 indeterminate positives prepared and preserved in the testing laboratory were tested. Serum samples, the test results are as follows:

[0063] Table 8 The results of negative and positive samples detected by indirect hemagglutination reagent of multiple serotypes of Haemophilus parasuis

[0064] Type 1

Type 2

type 3

Type 4

Type 5

...

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Abstract

The invention relates to the field of veterinary detection reagents, and discloses a haemophilus parasuis indirect hamagglutination detection reagent. The haemophilus parasuis indirect hamagglutination detection reagent is prepared by taking multiple serotype haemophilus parasuis whole protein mixtures as antigens, and is capable of rapidly and sensitively detecting serum antibodies of all serotype haemophilus parasuis diseases. The production technology is optimized through balanced mixing of each whole protein antigen, and then sensibilization and double hydroformylation of mutton red cells; and the prepared haemophilus parasuis indirect hamagglutination detection reagent is stable, sensitive, good in specificity, long in storage life, simple in operation, and applicable to a large amount of clinical sample detection on the haemophilus parasuis diseases, immune level detection and epidemiology investigation.

Description

technical field [0001] The invention relates to the field of veterinary detection reagents, in particular to an indirect hemagglutination detection reagent for Haemophilus parasuis, and also relates to a preparation method of the reagent. Background technique [0002] Haemophilus parasuis (Hps) is a Gram-negative coccus bacillus in the genus Haemophilus of the Pasteurella family. It is an opportunistic pathogen of pigs and is distributed worldwide. Porcine Glasser's disease characterized by polyserositis, arthritis and meningitis. It mainly harms piglets aged 2 to 8 weeks, the incidence rate is generally 10% to 15%, and the mortality rate can reach more than 50%. [0003] There are great differences in antigenicity and virulence among Hps strains, and serological typing is one of the important methods to distinguish different strains. At present, the serotyping method (KRG) based on the thermostable antigen immunodiffusion test proposed by Kielstein et al. in 1992 has been...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
Inventor 王占伟刘茂军邵国青冯志新熊祺琰白方方王海燕白昀吴叙苏王丽
Owner JIANGSU ACAD OF AGRI SCI
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