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Diagnostic value of fluorescence PCR melting curve method for drug resistance of Isoniazid, Ethambutol, and Levofloxacin against tuberculosis#br#
SHI Jianquan1   GAO Shan2   WANG Jun2
1.Department of Critical Care Medicine, Beijing Chest Hospital, Capital Medical University, Beijing   101149, China; 
2.Department of Internal Medicine-Tuberculosis, Beijing Chest Hospital, Capital Medical University, Beijing   101149, China
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Abstract  Objective To evaluate the diagnostic value of fluorescence PCR melting curve method for drug resistance of Isoniazid, Ethambutol, and Levofloxacin against tuberculosis. Methods A retrospective analysis was performed on 170 tuberculosis patients admitted to Beijing Chest Hospital, Capital Medical University from January 2018 to December 2019. A comparison was made between phenotypic drug susceptibility testing (DST) and fluorescence PCR melting curve method for anti-tuberculosis drug resistance. The diagnostic value of fluorescence PCR melting curve method for anti-tuberculosis drug resistance was analyzed using phenotypic DST results as the gold standard. The evaluation indexes included sensitivity, specificity, coincidence rate, Kappa value, and area under the curve (AUC) value. The drug resistance of quinolones (Levofloxacin and Moxifloxacin) was further compared. Results The number of cases of DST sensitivity by fluorescence PCR melting curve method and phenotype DST resistance was 5 cases in Isoniazid, 5 cases in Ethambutol, and 3 cases in Leofloxacin. The number of cases with DST resistance by fluorescence PCR melting curve method and phenotype DST sensitive was 9 cases in Isoniazid, 16 cases in Ethambutol, and 24 cases in Leofloxacin. The overall inconsistent rates of Isoniazid, Ethambutol, and Levofloxacin were 8.24%, 12.35%, and 15.88%, respectively. The sensitivity, specificity, coincidence rate, Kappa value, and AUC value of fluorescence PCR melting curve method to detect Isoniazid resistance were 91.53%, 91.89%, 91.76%, 0.821, and 0.917, respectively; the sensitivity, specificity, coincidence rate, Kappa value, and AUC value of fluorescence PCR melting curve method to detect Ethambutol resistance were 77.27%, 89.19%, 87.65%, 0.548, and 0.832, respectively; the sensitivity, specificity, coincidence rate, Kappa value, and AUC value of fluorescence PCR melting curve method to detect Levofloxacin resistance were 89.66%, 82.98%, 84.12%, 0.564, and 0.863, respectively. The drug resistance rate of Moxifloxacin against Mycobacterium tuberculosis (2.94%) was significantly lower than that of Levofloxacin (17.06%) (P < 0.01). Conclusion Fluorescence PCR melting curve method has good auxiliary diagnostic value for drug resistance of Isoniazid, Ethambutol, and Levofloxacin.
Key wordsFluorescence PCR melting curve      Drug susceptibility testing      Tuberculosis      Drug resistance      Synonymous mutation      Heterogeneous drug resistance     
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SHI Jianquan1 GAO Shan2 WANG Jun2
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SHI Jianquan1 GAO Shan2 WANG Jun2. Diagnostic value of fluorescence PCR melting curve method for drug resistance of Isoniazid, Ethambutol, and Levofloxacin against tuberculosis#br#[J]. 中国医药导报, 2022, 19(4): 37-40,44.
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