Nested PCR (polymerase chain reaction) primer group for simultaneously detecting mycoplasma hyopneumoniae and swine chlamydiosis and application of nested PCR primer group

Abstract

The invention provides a nested PCR primer group for simultaneously detecting mycoplasma hyopneumoniae and swine chlamydiosis and application of the nested PCR primer group, and belongs to the technical field of molecular diagnosis. The nested PCR primer group for simultaneously detecting the mycoplasma hyopneumoniae and the swine chlamydiosis, provided by the invention, comprises two pairs of outer side primers including Mhp-O-F and Mhp-O-R as well as C.suis-O-F and C.suis-O-R and two pairs of inner side primers including Mhp-I-F and Mhp-I-R as well as C.suis-I-F and C.suis-I-R. According to the primer group, an Mhp M12916S rDNA gene sequence and a C.suis ompA gene conserved sequence are taken as diagnostic targets for detecting mycoplasma hyopneumoniae and swine chlamydiosis, two pairs of outer primers and two pairs of inner primers of nested PCR are designed, mycoplasma hyopneumoniae and swine chlamydiosis can be detected at the same time, two rounds of PCR improve the sensitivity, anti-interference performance and specificity of PCR, and the defects that common PCR detection is low in sensitivity, poor in specificity and the like are overcome; and compared with the existing indirect immunofluorescence, serology and other detection methods, the cost is relatively low, and the detection period is short.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a nested PCR primer set for simultaneously detecting mycoplasma hyopneumoniae and chlamydia swine and its application. Background technique [0002] Mycoplasma hyopniumoniae (Mhp) is the pathogenic microorganism that causes Mycoplasma pneumonia of Swine (MPS). Mycoplasma suis pneumonia is widely prevalent in various pig raising areas all over the world. The increase in medical costs and the decline in pig production performance caused by the prevalence of the disease have caused huge losses to the pig industry every year. Mycoplasma hyopneumoniae is one of the main pathogens of porcine respiratory syndrome (porcine respiratory disease complex, PRDC) besides directly causing the loss of mycoplasma pneumoniae. [0003] Swine Chlamydiosis (C.suis) is a chronic contact infectious disease caused by certain strains of Chlamydia psittaci (formerly known as Chlam...

AI Technical Summary

Problems solved by technology

The isolation and culture of pathogens is the gold standard for the diagnosis of all pathogenic microorganisms including mycoplasma, but in most cases it will not be used as a laboratory method for the diagnosis of Mycoplasma hyopneumoniae and Chlamydia swine infection; mainly because of the inherent difficulty and slowness of mycoplasma isolation

In the laboratory, immunofluorescence is often used to detect pathogens, but this method has the disadvantage of insufficient sensitivity

Serological diagnostic methods can be applied to detect whether the population is infected with Mycoplasma hyopneumoniae and Chlamydia suis, but this method is not suitable for individuals, because individuals infected with Mycoplasma hyopneumoniae and Chlamydia suis have different virulence strains, individual health status, Under the influence of multiple factors such as the maternal antibody level, the time for the antibody to become positive varies from a few weeks to a few months

Polymerase chain reaction (Polymerase chain reaction, PCR), as one of the commonly used laboratory diagnostic methods for detecting Mycoplasma hyopneumoniae and Chlamydia swis, has the characteristics of rapidity. Cross-infection will occur at the same time as positive

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