Hepatic abscess-causing Klebsiella pneumoniae specific gene sequence lak and its application
A technology for Klebsiella pneumoniae and Klebsiella pneumoniae liver, which is applied in the field of medical disease diagnosis and treatment drug selection, can solve the problems of easy missed diagnosis, delayed diagnosis of Klebsiella pneumoniae liver abscess, and low culture positive rate, etc. Highly specific effect
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Embodiment 1
[0034] The screening process of specific gene sequences combines the next-generation sequencing technology (Next-Generation Sequencing, NGS), comparative genomics (Comparative Genomics) and bioinformatics (Bioinformatics) methods, and uses scientific statistical methods to analyze sequences in multiple The specificity of the distribution in the Klebsiella pneumoniae genome was tested, and PCR validation was performed in the isolated Klebsiella pneumoniae and non-hepatoabscess Klebsiella pneumoniae to determine the sequence in Klebsiella pneumoniae. Specificity in Lebsiella. The specific implementation process is as follows:
[0035] Pathogen isolation and identification:
[0036] 1. Enrollment of patients with suspected Klebsiella pneumoniae liver abscess. Patients with clinically suspected Klebsiella pneumoniae liver abscess were screened according to the following criteria:
[0037] (1) Clinical manifestations of the patient: fever, chills and abdominal pain;
[0038] (2...
Embodiment 2
[0061]Gene-specific verification:
[0062] The specific steps for the verification of three hepatic abscess-causing Klebsiella pneumoniae-specific genes in clinically isolated strains are:
[0063] 1. Select clinically isolated hepatic abscess and non-hepatic abscess Klebsiella pneumoniae bacterial strains for detection of sdm (shown in SEQ ID NO.1), lak (shown in SEQ ID NO.3) and asnC (shown in SEQ ID NO.3) .2) specificity in hepatoabscess-causing Klebsiella pneumoniae. 17 strains of Klebsiella pneumoniae isolated from the abscess of patients with liver abscess were selected as the detection group (hepatic abscess group), and 17 strains of Klebsiella pneumoniae isolated from non-Klebsiella pneumoniae patients with liver abscess were selected as negative controls group (non-hepatic abscess-induced group). At the same time, other common pathogenic bacteria (other pathogenic bacteria groups) isolated clinically were also selected to further verify the specificity of these fact...
Embodiment 3
[0086] Application of sdm, lak and asnC sequences or primers designed based on their sequences in the preparation of Klebsiella pneumoniae liver abscess diagnostic kit:
[0087] The design idea of this experiment is as follows: the clinically obtained blood culture samples are divided into a blood culture positive group and a healthy person negative group, where the blood culture positive group refers to all samples with bacteria cultured in the blood, not limited to the blood cultured with pneumonia. Lebsiella, including cultures of other pathogenic bacteria. Use PCR to detect the presence of specific factors (i.e. sdm, lak and asnC) in positive blood culture samples and negative blood culture samples, and finally analyze the corresponding analysis of the PCR results and the patient's final clinical diagnosis, so as to judge whether the method provided by the present invention is suitable for Detection rate of Klebsiella pneumoniae liver abscess.
[0088] 1. Collection of ...
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