A multiple fluorescence immunoassay method and reagents for rapidly distinguishing aiv, ndv, mg and ms

Abstract

The invention discloses a multiple fluorescent immunoassay method and reagent for quick distinguishing of avian influenza virus, Newcastle disease virus, Mycoplasma gallisepticum and Mycoplasma synoviae. The method is simple to perform, a target amplification fragment is acquired through PCR (polymerase chain reaction), an amplification product, fluorescence coded microspheres and streptavidin-phycoerythrin are hybridized, and MFI (mean fluorescence intensity) value is read through a detector to distinguish different types of pathogens. The method enables accurate detection for avian influenza virus, Newcastle disease virus, Mycoplasma gallisepticum and Mycoplasma synoviae, and is high in specificity, high in sensitivity and good in repeatability; compared with traditional detection methods, the method implements simultaneous detection for various different target molecules in a same sample, fewer samples are used, the operation is simple, the operation speed is high, and detection cost can be greatly reduced; the method is good in flexibility and allows addition and deduction of the types of pathogen on such basis as required.

Description

technical field [0001] The invention belongs to the field of pathogen detection in the breeding industry, and in particular relates to a multiple fluorescence immunoassay method for rapidly distinguishing avian influenza (AIV), Newcastle disease virus (NDV), mycoplasma gallisepticum (MG) and mycoplasma gallisepticum (MS). 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 cannot be ignored in chicken production. ...

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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