A multiple fluorescence immunoassay method and reagents for rapidly distinguishing ilTV, ibv, mg and ms

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 the breeding industry, and specifically relates to a method for quickly distinguishing avian infectious laryngotracheitis virus (ILTV), avian infectious bronchitis virus (IBV), mycoplasma gallisepticum (MG) and mycoplasma synovialis ( MS) multiple fluorescence immunoassay methods and reagents. Background technique [0002] With the continuous increase in market demand for poultry, the poultry industry has developed rapidly, and the degree of intensification of the poultry industry has continued to increase, and the infection of avian respiratory pathogens has been on the rise year by year. Poultry respiratory diseases have always been the main factor leading to significant economic losses in the poultry industry. The pathogens of chicken respiratory diseases are complicated. It can be caused by viruses, mycoplasma and bacteria, and various mixed infections will appear in the clinic. The occurrenc...

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

Images