Multiplex quantitative PCR kit for rapid joint detection of four difficult-to-culture and identification bacteria

Abstract

The invention provides a multiple quantitative PCR (polymerase chain reaction) kit for quick combined detection of four bacteria difficult to cultivate and identify. The multiple quantitative PCR kit comprises four PCR reaction systems, wherein the four PCR reaction systems comprise AllGlo fluorescence probes and forward and reverse primers aimed at the following four pathogens which cause child bacterial pneumonia and are clinically difficult to cultivate and identify: haemophilus influenza, streptococcus pneumonia, moraxella catarrhalis and legionella pneumophila. According to the multiple quantitative PCR kit, the design is reasonable, the infection of the four pathogens, clinically difficult to cultivate and identify, of child bacterial pneumonia can be easily, conveniently, quickly and parallelly detected in a reaction tube at the same time, the situation that four bacteria are detected at the same time through single-tube PCR is achieved, quantitative detection is achieved, the kit is easy and quick to operate, high in sensitivity, good in specificity and repeatability, accurate and reliable in result, early specific diagnosis, prevention and treatment can be provided for patients suffering from infantile pneumonia according to the bacterial infection titer, and the kit has great clinical practicability for interdicting an infection source, reducing infection or mixed infection of the four bacteria and monitoring the clinical curative effect.

Description

technical field [0001] The invention belongs to biotechnology, and relates to a multiplex quantitative PCR kit for rapid joint detection of four difficult-to-culture and identification bacteria, in particular to the detection of Haemophilus influenzae (Haemophilus Influenzae, HI), Streptococcus pneumoniae (Streptococcus Pneumoniae, SP), catarrhal Quadruple quantitative PCR kits for Moraxella (Moraxella Catarrhalis, MC) and Legionella Pneumophila (Legionella Pneumophila, LP). Specifically, the present invention relates to the application of quadruple quantitative PCR technology in a PCR reaction tube to rapidly and parallel A kit for simultaneously detecting four types of bacteria (HI, SP, MC, LP). Background technique [0002] According to the World Health Organization (WHO), pneumonia is the "number one killer" of children worldwide and the second cause of death for children under 5 years of age. In my country, childhood pneumonia is also an important common disease in chi...

AI Technical Summary

Problems solved by technology

Sputum bacterial culture is still a routine test item. Positive bacterial culture is the gold standard for diagnosis and has certain clinical value. However, it grows slowly, takes a long time to culture, and has high technical requirements. It is difficult to isolate and culture from clinical samples and difficult to identify. Easy to confuse and cause misdiagnosis, especially in mixed infections, some potential pathogenic bacteria may be overlooked, thus limiting its clinical application

In addition, although the serological method is mature and commercial kits are available, the sensitivity and specificity need to be improved. At the same time, it is necessary to detect double serum in the acute phase and the convalescent phase to be diagnostically meaningful and it is easy to miss the diagnosis.

Images