Loop-mediated isothermal amplification detection method of E. wenyoni

Abstract

The invention belongs to the technical field of detection of parasite molecules, and in particular relates to a quick loop-mediated isothermal amplification detection method of E. wenyoni, which comprises the following steps: (1) designing three pairs of specific primers; (2) extracting total DNA of a blood sample; and (3) carrying out loop-mediated isothermal amplification reaction and analyzing the amplification product. By imaging on a gel imaging system and observing the result, E. wenyoni is positive if the amplification product is a stepped pattern formed by zones of different sizes; or SYBR Green I is added to the final product of amplification, and by visually observing the result under natural light, the E. wenyoni is positive if the amplification product turns to green yellow, and the E. wenyoni is negative if the amplification product is reddish brown. The invention has strong specificity and can be used for discriminating common blood protozoa, the sensitivity is high and is 10-100 times higher than that of common PCR, the detection is simple and quick, only one water bath is needed, and the result can be obtained in 1 hour.

Description

technical field [0001] The invention belongs to the technical field of parasite molecular detection, and in particular relates to the application of a loop-mediated isothermal amplification method in the in vitro detection of Eperythrozoon wennerii. technical background [0002] Eperythrozoonosis is caused by the obligate blood parasite Eperythrozoon, and is a disease characterized by anemia, jaundice, and fever. Some people at home and abroad have called it icteric anemia, anaminidosis, red animal disease and erythroderma, etc. 【1】 . Since Schilling and Dingen discovered the sphagoid haemozoa in rodents in 1928, scientists have successively discovered the existence of the sphagoid haemozoa in different animals. Among them, in 1934, Adler et al. discovered a microorganism similar in shape to the spheroid hematocides in cattle, and named it E. wenyoni. [0003] The classification of Eperythrozoon has always been controversial. In 1974, the eighth edition of "Berger's Bac...

AI Technical Summary

Problems solved by technology

[0004] PCR amplification of the 16S rRNA gene of Eperythrozoon bovis at the molecular level can identify pathogens in a timely and accurate manner, but ordinary PCR and more sensitive Nested PCR and fluorescent quantitative PCR must rely on expensive precision instruments, high testing costs, high technical requirements for testing personnel, etc., and cannot be carried out in basic laboratories with poor conditions

Images