Double nested PCR (polymerase chain reaction) method capable of simultaneously detecting trichomonas hominis and trichomonas brateri and application of double nested PCR method

Abstract

The invention belongs to the technical field of molecular detection, and relates to a double nested PCR (polymerase chain reaction) method capable of simultaneously detecting trichomonas hominis and trichomonad of cattle and application of the double nested PCR method. Amplification is performed by taking a trichomonas hominis and trichomonad positive control mixture as a template and trichomonad specific primers as amplification primers; an amplification product obtained by the first round of PCR is used as a template, a mixture of a trichomonas hominis species specific primer and a trichomonas brateri species specific primer is used as a primer for amplification, and the trichomonas hominis and the trichomonas brateri are detected at the same time. The method disclosed by the invention can specifically amplify target fragments of trichomonas hominis and trichomonas brateri, and can detect positive control DNA (Deoxyribonucleic Acid) of 500fg / mu L to the minimum.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and relates to a double nested PCR method capable of simultaneously detecting Pentatrichomonas bovis and Tetratrichomonas batellii and its application. Background technique [0002] Trichomonas is a protozoa that parasitizes the digestive system and urogenital tract of vertebrates. It is the oldest eukaryote, including both pathogenic and some symbiotic species. Trichomonas is also a very common protozoan in cattle. The currently reported species of Trichomonas mainly include Tritrichomonas foetus (T.foetus), and some species of Trichomonas that parasitize the intestinal tract of cattle. Such as Pentatrichomonas hominis (P.hominis), Tetratrichomonas buttreyi (T.buttreyi) and so on. Among them, Trichomonas fetalis is the most common Trichomonas bovis, which parasitizes in the reproductive organs of cows and bulls, and can cause a sexually transmitted protozoan disease of cattle, namel...

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Problems solved by technology

In addition, light microscopy and culture methods require that the samples used, such as feces or bovine genital tract washings, must be fresh, and there are disadvantages such as false negatives due to refrigerated or failure to process samples in time to cause parasite death.

At present, some molecular methods for the detection of Tritrichomonas fetalis have been developed, but there are few reports on the molecular methods for the detection of Pentatrichomonas hominis and Tetratrichomonas batellii

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