Orientia tsutsugamushi nucleic acid fluorescence isothermal amplification primer, probe, kit and detection method

Abstract

The invention provides an orientia tsutsugamushi nucleic acid fluorescence isothermal amplification primer. The primer is screened from an Ot1 sequence, the primer comprises an upstream primer and a downstream primer, the upstream primer has the sequence characteristic of SEQ ID NO.1, and the downstream primer has the sequence characteristic of SEQ ID NO.2. The invention provides an orientia tsutsugamushi nucleic acid fluorescence isothermal amplification probe, the probe is screened from an Ot1 sequence, and the probe (Ot-P) has the sequence characteristic of SEQ ID NO.3. The invention discloses an orientia tsutsugamushi nucleic acid fluorescence isothermal amplification kit. The kit comprises the primer and the probe. The invention provides a detection method using the orientia tsutsugamushi nucleic acid fluorescence isothermal amplification kit. The detection method comprises the following steps: S1, treating a template; s2, treating the template, the freeze-dried enzyme powder, the primer, the probe, ultrapure water (Ultrapure water, UP water, the resistivity of which reaches 18 M omega.cm) and an activating agent, and reacting at the temperature of 35-41 DEG C for 20-40 minutes. The primer and the probe are high in specificity and sensitivity; the detection time is short, large expensive instruments and variable temperature systems are not needed, nucleic acid amplification can be completed at 37 DEG C within 30 minutes, and on-site rapid nucleic acid detection and popularization are easy.

Description

technical field [0001] The invention relates to the field of molecular biology detection of Orientia tsutsugamushi, in particular to a fluorescent isothermal amplification primer, probe, kit and detection method of Orientia tsutsugamushi nucleic acid. Background technique [0002] Scrub typhus, also known as scrub typhus, is a zoonotic infectious disease transmitted by Orientia tsutsugamushi (Ot) through the bites of chigger larvae. The main clinical manifestations of scrub typhus patients are fever, rash, eschar or ulcer, headache, muscle aches, and swollen lymph nodes. Studies have reported that about 1 million people in the world are infected with scrub typhus every year, and nearly 1 billion people are at risk of being infected with scrub typhus. From 2006 to 2016, the number and incidence of scrub typhus in my country increased year by year, mostly in southern China. Scrub typhus is similar to other febrile diseases in terms of symptoms and signs. It is often misdiagn...

AI Technical Summary

Problems solved by technology

Although PCR detection is sensitive, its high detection cost, the need for specific equipment, professional technicians and other defects limit its wide practical application

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