Multiple fluoroimmunoassay method capable of quickly differentiating infectious laryngotracheitis virus (ILTV), infectious bronchitis virus (IBV), myeoplasma gallisepticum (MG) and mycoplasma synoviae (MS) and reagent

Abstract

The invention discloses a multiple fluoroimmunoassay method capable of quickly differentiating infectious laryngotracheitis virus (ILTV), infectious bronchitis virus (IBV), myeoplasma gallisepticum (MG) and mycoplasma synoviae (MS) and a reagent. The method is simple to operate, a target amplified fragment is obtained through polymerase chain reaction (PCR); an amplified product, fluorescence coded microspheres and streptavidin-phycoerythrin are hybridized, a mean fluorescence intensity (MFI) value is read through a detector, and pathogens of different types are differentiated. According to the method, avian infectious laryngotracheitis virus, infectious bronchitis virus, myeoplasma gallisepticum and mycoplasma synoviae can be accurately detected simultaneously, and the method is high in specificity and sensitivity and good in repeatability. Compared with a conventional detection method, the method can be used to simultaneously detect various target molecules in an identical sample, the sample usage amount is small, the method is simple and quick to operate, and the detection cost can be greatly reduced. The method is good in flexibility, and the varieties of detected pathogens can be increased or decreased as required on the basis.

Description

technical field [0001] The invention belongs to the field of pathogen detection in aquaculture, in particular to a method for rapidly distinguishing chicken infectious laryngotracheitis virus (ILTV), chicken infectious bronchitis virus (IBV), mycoplasma gallisepticum (MG) and mycoplasma gallisepticum ( MS) multiple fluorescence immunoassay method and reagents. Background technique [0002] As the market demand for poultry continues to increase, the poultry industry has developed rapidly, the degree of intensification of the poultry industry has continued to increase, and the infection of poultry respiratory pathogens has been on the rise year by year. Respiratory diseases of poultry have been a major factor causing significant economic losses in the poultry industry. The pathogenesis of chicken respiratory disease has its complexity. It can be caused by viruses, mycoplasma and bacteria, and various mixed infections will appear clinically. The occurrence of these diseases c...

AI Technical Summary

Problems solved by technology

Compared with conventional PCR, fluorescent quantitative PCR has advantages in terms of sensitivity, specificity, and speed. However, in practical applications, when the sample volume is very large, single-plex fluorescent PCR has certain disadvantages in terms of cost and time. A multiplex fluorescent PCR method is much more complicated than a single-plex one, and it has higher requirements on reagents and primers. At the same time, it is necessary to ensure that there is no mutual interference between the fluorescent groups labeled by different probes. The fluorescent quantitative PCR instrument used has corresponding Multiple detection channels make the experiment more difficult

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