Primer probe and kit for detecting lung pathogenic bacteria

Abstract

The invention relates to the technical field of biological detection, in particular to a primer probe combination for detecting lung pathogenic bacteria, application of the primer probe combination and a kit. The primer probe combination disclosed by the invention can realize rapid detection of four pulmonary infection bacteria, namely aspergillus, Yarrowia, cryptococcus and nocardia, is good in accuracy and specificity and high in sensitivity, facilitates real-time evaluation of the health condition of a patient, timely provides adjuvant therapy suggestions for clinicians, and has important significance for improving the survival rate of infected patients.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer-probe combination and a kit for detecting pulmonary pathogenic bacteria. Background technique [0002] Pulmonary infection refers to inflammation of the lung parenchyma including the terminal airways, alveolar space and pulmonary interstitium. Pulmonary infection includes bacterial lung infection, fungal lung infection, viral lung infection, mycoplasma lung infection, etc. Because of their wide variety, this project focuses on invasive fungal infections of the lung and Nocardia spp. Pulmonary erosive fungi also include: Candida, Aspergillus, Cryptococcus, Mucor and Pneumocystis in the phylum Zygomycetes. Because Candida is mainly infected through the bloodstream, the incidence of pulmonary infection in the phylum Zygobacter is 2.9 / 100,000 and the mortality rate is 86%. It is very difficult to obtain samples. The lung invasive fungi that this project mainly...

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Problems solved by technology

[0008] These detection methods have their own advantages and disadvantages. Smear microscopy is easy to operate, but the possibility of infection in negative patients cannot be ruled out.

Although histopathological examination is the gold standard for deep infection detection, it cannot determine the pathogenic bacteria species

Bacterial culture is the isolation of opportunistic pathogenic bacteria from sterile parts such as blood, pleural effusion, bronchoalveolar lavage fluid, and biopsy tissue blocks, which often indicates a definite infection, but the results should be interpreted cautiously for sputum and other specimens. Positive results often cannot confirm the diagnosis, and repeated inspections should be repeated if necessary. At the same time, negative results cannot rule out invasive aspergillosis. Drugs should be selected based on drug sensitivity. Unable to cultivate, easy to lose the best treatment time

The detection of serum antigens and their metabolites includes GM test and G test: GM test has a sensitivity of 80.7% and a specificity of 89.2% in the diagnosis of invasive aspergillosis. Although it can be used as a basis for the diagnosis of invasive aspergillosis, it is susceptible to certain The influence of some foods or drugs can cause false positive results, and the false positive rate is as high as 18%. Although the G test is included in the diagnostic criteria for invasive fungal diseases, it cannot be used for the detection of zygomycete and cryptococcal infection, and it cannot be differentiated Pathogenic species with a high false positive rate

Although fluorescent quantitative PCR and loop-mediated technology are time-consuming and high-efficiency, limited by the second-generation PCR technology itself, its detection depth can only reach 1%, and its sensitivity is low

[0009] Therefore, the current clinical lung infection detection technology not only has problems such as long time-consuming, poor specificity, high false positive rate and missed detection rate, poor repeatability, etc., but also mainly stays at the level of qualitative judgment. There is still a long way to go to meet clinical needs in accurately distinguishing bacterial species and other issues

However, there is no report on the application of digital PCR to the combined detection of different genera of lung pathogenic bacteria.

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