Primer, probe and kit for detecting and typing rickettsia by virtue of real-time FRET-PCR (Fluorescent Resonance Energy Transfer-Polymerase Chain Reaction) and nested PCR

Abstract

The invention relates to a primer, a probe and a kit for detecting and typing a rickettsia by virtue of real-time FRET-PCR (Fluorescent Resonance Energy Transfer-Polymerase Chain Reaction) and nested PCR. The primer and the probe are shown as sequences 1-8. By virtue of designing the primer and the probe of the FRET-PCR, nucleinic acids of all Rickettsiaes can be specifically amplified, thus fast judging positive samples; then two pairs of primers of the nested PCR can be designed by selecting other conservative intervals of the same gene, and the Rickettsiae of the corresponding positive samples can be determined by virtue of nested PCR, agarose gel electrophoresis and sequencing. Therefore, the system not only can fast and specifically detect the rickettsia organisms of all the Rickettsiaes, but also can confirm the Rickettsiae.

Description

technical field [0001] The invention establishes a system based on citrate synthase gene, which can detect and type all Rickettsia species. The system achieves the purpose of rapid detection and typing of all Rickettsia species through the combination of real-time FRET-PCR and nested PCR. Background technique [0002] Rickettsiae are bacteria, but at the same time have the characteristics of viruses, are strictly intracellular parasites, parasitic in various blood-sucking arthropods and insects. Humans and animals are infected by being bitten by or touching the feces of Rickettsia-infected vector insects, which can cause clinical symptoms such as fever, headache and rash in humans and animals. The disease mostly occurs in tropical and subtropical countries and regions, and Rickettsia infection has become a worldwide problem at present. At present, my country has listed epidemic typhus, endemic typhus and other infectious diseases caused by Rickettsia infection as Class B i...

AI Technical Summary

Problems solved by technology

However, current molecular diagnostic methods are not mature enough

Some detection methods through traditional PCR and agarose gel electrophoresis can cover all Rickettsia species, but the operation is cumbersome and not suitable for the detection of a large number of samples

Existing quantitative PCR can only detect some species of Rickettsia, and cannot distinguish different species of Rickettsia with differences in pathogenicity

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