Method and kit for identifying and detecting babesia unidentified species

Abstract

The present invention discloses a cross-primer constant temperature amplification method and a kit used for identifying and detecting babesia unidentified species. The constant temperature amplification kit for identifying and detecting the babesia unidentified species comprises a cross primer BXJ-2a1s having a sequence of SEQ ID No.3, a replacement primer BXJ-5a having a sequence of SEQ ID No.1 and a replacement primer BXJ-4s having a sequence of SEQ ID No.2, respectively, a probe BXJ-2a having a sequence of SEQ ID No.4 and a probe BXJ-3a having a sequence of SEQ ID No.5, respectively, conventional reagents for amplification, and lateral flow chromatography test strips required for determination of results. The detection method can specifically detect the babesia unidentified species, isfast, simple in operation, strong in specificity, high in sensitivity, free of complicated instruments and equipment, and intuitive and visible in results, and very suitable for rapid detection at grass-root sites and has relatively large application prospects.

Description

technical field [0001] The present invention relates to a method and a kit for detection and differential detection of Babesia unspecified species by means of constant temperature amplification of cross primers combined with immunochromatographic technology. Specifically, the present invention relates to a detection kit for Babesia unspecified species and non-disease diagnosis method. Background technique [0002] Ovis babesiosis of sheep is a blood protozoa characterized by fever, jaundice, hemoglobinuria, etc. The disease, which is transmitted by ticks, can cause death in severe cases (Uilenberg, 2006). The pathogens causing sheep babesiosis reported so far in the world are Babesia ovis, Babesia motasi, Babesia crassa, Babesia foliata ) and Babesia taylori (Babesiataylori), the pathogenicity of which weakened in turn. Epidemiological surveys and studies on the biological characteristics of the disease have shown that there are two species of Babesia prevalent in my coun...

AI Technical Summary

Problems solved by technology

[0003] At present, the classic blood smear staining method is used to detect the infection of Babesia sheep. This method is difficult to detect the pathogen when the infection rate is very low, and the missed detection rate is high. Moreover, the morphologically A kind of identification and higher skill requirements for operators

Therefore, parasitologists have developed a series of Babesia identification and detection methods based on molecular biology, including real-time fluorescent quantitative PCR, reverse linear blotting, multiplex PCR, etc. These methods are characterized by high sensitivity and specificity, but Real-time fluorescent quantitative PCR and multiplex PCR require expensive instruments, and multiplex PCR also requires gel electrophoresis analysis of the amplified products, which requires high skill of the operator; reverse linear blot also requires pre-amplification of the sample to be tested, which is cumbersome to operate , time-consuming, and not suitable for the detection of batch samples, the detection throughput is low

Moreover, the above method cannot meet the application of on-site rapid detection.

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