Real-time PCR specific primer of hepatozoon, as well as probe, detection kit and detection method thereof

Abstract

The invention provides a real-time PCR specific primer of hepatozoon, as well as probe, a detection kit and a detection method thereof. According to the specific primer, the 18s rRNA gene of hepatozoon is used as a target gene, which has a highly conserved sequence, PCR can be easily amplified, two specific primers and a probe are designed by combining with related information of 18sr RNA genes ofother sporozoans for the specific region taking the 18srRNA of Hepatozoon spp. as the target gene according to the basic principle of TaqMan real-time fluorescent PCR, a simple, convenient, efficientand practical DNA detection method is established, and microgenome DNA extracted from affected dog blood can be directly used as a template for identifying. The method can be used for quickly detecting hepatozoon diseases, has a wide application prospect, and can accurately and quickly determine whether a cow is affected by hepatozoon or not.

Description

technical field [0001] The invention specifically relates to the technical field of biology, in particular to a Real-time PCR specific primer and probe, a detection kit and a Real-time PCR detection method for liver clusterworms. Background technique [0002] Liver cluster canine is a sporozoite that reproduces asexually in dogs and sexually in ticks. The worm is widely distributed in Africa, southern Europe, the Middle East and Asia, and its host range is also wide, and various vertebrates can be infected. Dogs infected with hepatic clusterworm generally manifest as anorexia, weight loss, lack of energy, occasional vomiting and diarrhea and other gastrointestinal symptoms; sometimes unsteady gait and abnormal gait; Pale, painful response to palpation, occasional subcutaneous hemorrhage, gingival bleeding and abdominal purpura; when the infection is severe, the body becomes hot, and when the schizont invades the viscera, the corresponding function is destroyed; if the kidne...

AI Technical Summary

Problems solved by technology

The pathogenic detection methods of hepatic fasciculosis mainly focus on blood smear and muscle section microscopic examination, but the efficiency of blood smear and muscle section microscopic examination is low and cannot be used as a reliable detection method

Immunological detection is mainly an indirect ELISA method, but this type of method cannot distinguish whether the animal is being infected or has been infected, and cannot identify the carrier animal and the storage host

PCR technology is one of the most widely used methods available for the diagnosis of hepatic fasciculosis. Conventional PCR detection technology has a big disadvantage, low specificity, and false positives are easy to occur during amplification. Cross-reactivity between hepatic clusterworms and other sporozoites was found in the process of

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