Nucleic acid typing detection kit for encephalitis and meningitis and detection method

Abstract

The invention discloses a nucleic acid typing detection kit for encephalitis and meningitis. The kit comprises a nucleic acid rapid extraction reagent, a PCR amplification reagent, an encephalitis andmeningitis nucleic acid detection reagent, a positive reference substance and a negative reference substance. The encephalitis and meningitis nucleic acid detection reagent comprises an amplificationprimer and a probe of a streptococcus pneumoniae gene, an amplification primer and a probe of a neisseria meningitidis gene, an amplification primer and a probe of a haemophilus influenzae gene, andan amplification primer and a probe of an internal reference RNP gene. The invention further provides a nucleic acid typing detection method for encephalitis and meningitis. According to the invention, multi-channel primers and probes with encephalitis and meningitis specificity are introduced; the workload can be reduced; the problem that other detection methods are low in specificity and prone to missed diagnosis and misdiagnosis is solved; multiple simultaneous detection of three pathogens in the same system can be achieved, and primers and probes in all channels do not interfere with one another; the sensitivity is good; the specificity is high; accuracy and reliability are achieved; rapidness and convenience are achieved; and whether the three pathogens are infected or not is rapidlydiagnosed.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to an RT-PCR amplification technology, in particular to an encephalitis meningitis nucleic acid typing detection kit and a detection method. Background technique [0002] Meningitis and encephalitis are a type of syndromes that can be caused by bacteria, viruses, and yeasts that threaten the lives of children. Morbidity and mortality are high, especially bacterial fungal meningitis. The incidence of acute bacterial meningitis in children under five years of age in my country is (6.95~22.3) / 10,000. Meningitis and encephalitis are caused by infection of the central nervous system. After 40% of patients are infected, the course of the disease progresses. Faster, directly leading to death, and survivors will also be left with neurological defects. Encephalitis and meningitis may exist at the same time, become encephalitis meningitis, or have the same symptoms, such as headache and fever. Accordi...

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Problems solved by technology

[0008] Chinese patent document CN201610296666.5 discloses a multiplex PCR kit for detecting bacterial meningitis pathogens, which can rapidly detect six kinds of bacterial meningitis pathogens, Streptococcus pneumoniae (SP), Streptococcus agalactiae (GBS), influenzae Haemobacterium (HI), Listeria monocytogenes (LM), Staphylococcus aureus (SA), Neisseria meningitidis (NM), the detection method is a multiplex PCR method, and the amplification results are detected by electrophoresis; Need to open the cover for detection, which may bring potential contamination, and the method is complicated to operate, and the detection sensitivity is lower than that of fluorescent quantitative PCR method

[0009] Chinese patent document CN201310031419.9 discloses a kit for synchronously detecting 23 pathogens of encephalitis and meningitis and its detection method, which can detect Neisseria meningitidis, Haemophilus influenzae type b, Staphylococcus, Streptococcus pneumoniae For pathogens including cocci, etc., the detection method is capillary electrophoresis. Because the product needs to be opened for detection, it may cause potential contamination, and the operation is more cumbersome, the detection time is long, and the detection sensitivity is lower than that of fluorescent quantitative PCR method.

[0010] Chinese patent document CN201110004826.1 discloses a nucleotide specific to ITS of Neisseria meningitidis and its application. The fluorescent quantitative PCR kit is used to detect Neisseria meningitidis in the human body and the environment. It can be used in the supervision and testing of food and clinical samples, the detection of pathogenic bacteria in food, bacteriological classification and epidemiological investigation, etc., but its main shortcoming is that the target is too single when testing samples

[0011] Although there are many pathogen detection kits for encephalitis and meningitis in the prior art, the triple detection method for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae has not been reported on the fluorescent quantitative PCR technology platform, so Therefore, it is urgent for those skilled in the art to use this platform to develop a high-sensitivity, high-specificity, and high-accuracy multiple real-time fluorescent quantitative PCR detection method and kit to realize triple joint detection and distinguish Streptococcus pneumoniae and Neisseria meningitidis Haemophilus influenzae

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