The Study Of Fluorescent Detection Nucleic Acid Analysy Technique Of Isothermal Amplification Microfluidic Chip For Rapid Strain Screening Of Suspectedjoint Tuberculosis

Objective:Through strains of suspected joint tuberculosiswere rapid screened by Fluorescent Detection Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip,and the feasibility,simplicity,shortcut and accuracy of the clinical application were introduced and evaluated.Methods:the 19 cases of suspected joint tuberculosis infection which admitted to the department and Chenggong branch of the First Affiliated Hospital of Kunming Medical University of orthopedics in April 2016-2017 October were selected.The medical history,clinical manifestations and imaging examination were considered as joint tuberculosis.Among them,there were 8 males and 11 females with an average age of 26.7 years,6 cases of joint replacement,13 cases without joint replacement,11 cases of tuberculosis of knee joint and 9 cases of tuberculosis infection of the hip joint.2.the 6ml was collected from the two groups of the same specimens of the joint puncture fluid,the tissue suspension in the joint replacement and the mixed fluid of the wound secretion in the patients with suspected joint tuberculosis infection.A group of specimens were immediately sent to the PCR room of our hospital to detect nucleic acid technology with Fluorescent Detection Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip(abbr.Isothermal Amplification Microfluidic,IAMC)method(the experimental group),and the other group was immediately sent to the bacterial culture room of infectious hospital of Kunming to be tested by Lowenstein-Jenden culture method(the control group).3.the detection time of Isothermal Amplification Technique and Lowenstein-Jenden culture was recorded,and the positive and negative numbers between the two groups were statistically analyzed.The results were obtained by comparing the data with the X2 test.Results:1.The results of Isothermal Amplification Technique(experimental group)were as follows:13 cases were positive,6 cases were negative,and Lowenstein-Jenden culture(control group)results were:7 cases were positive,12 cases were negative.Among them,3 specimens of non simple tuberculosis infection were detected in the experimental group:Staphylococcus aureus infection in 1 cases,joint Escherichia coli infection in 1 cases,joint tuberculosis with Staphylococcus aureus infection in 1 cases,3 specimens of the control group were negative.2.the Lowenstein-Jensen culture method was used to identify 7 Mycobacterium tuberculosis complex groups in the experimental group,12 cases were not detected tuberculosis bacteria,and the control group was not detected the Mycobacterium tuberculosis,so the sensitivity was 36.8%and the specificity was 100%.3.the Lowenstein-Jensen culture method time is about 4-8w,and the isothermal amplification time is about 180min.4.the positive rate of two kinds of tuberculosis detection methods was statistically significant(X2=5.115,P<0.05).The Kappa value is 0.424,and the consistency is good.Conclusion(s):1.Fluorescent Detection Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip can directly detect the sequence of specific Mycobacterium tuberculosis complex DNA amplified from the specimen,and quickly identify the bacteria to achieve the purpose of early diagnosis of tuberculosis infection.2.the sensitivity of Fluorescent Detection Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip was higher than that of Lowenstein-Jensen culture method,which could reduce the false negative of Lowenste in-Jensen culture method.3.the isothermal amplification technology does not require repeated thermal denaturation,such as denaturation,annealing,and so on.It is easy to operate and reduce the requirements of equipment and technology so as to facilitate the screening and detection of tuberculosis infection.4.in this study,the detection of nucleic acid by the Fluorescent Detection Nucleic Acid Analysy Technique of Isothermal Amplification Microfluidic Chip is superior to the other bacteria,which shows the good prospect of rapid screening for joint infection of joint.