Abstract: Objective To evaluate the application value of whole-genome sequencing technology to diagnostic the drug-resistant tuberculosis, and provide reference for the early diagnosis of drug-resistant tuberculosis. Method The phenotypic susceptibility results of 8 kinds of anti-tuberculosis drugs with sputum culture positive strains were regard as gold standard, which patients were confirmed and registered in the designated medical institution of Bao'an District from January 1, 2019 to December 31,2021. And the strains were inactivated before DNA was extracted for whole template sequence. The sequence results were uploaded to the platform to obtain the mutation sites of each strain. With reference to the drug resistance gene loci reported in previous literature, the sequence results were compared with the phenotypic drug susceptibility results, and the sensitivity, specificity,coincidence rate and Kappa value were used to evaluate the application value of whole-genome sequencing technology. Result The total drug resistance rate of 1327 culture-positive strains was 20.35%(270/1327). 86.30%(233/270) of the resistant strains were resistant to at least one of the four first-line anti-tuberculosis drugs, including isoniazid, rifampicin, ethambutol and streptomycin. The highest resistance rates of first and second-line anti-tuberculosis drugs were streptomycin(10.63%) and ofloxacin(3.01%). katG, fabG1,inhA and ahpC mutated genes were found in isoniazid-resistant strains. Three genetic mutations were found in resistant strains to streptomycin and para-aminosailcylic acid. Two genetic mutations were found in resistant strains to ethambutol, capreomycin and ofloxacin. There were only one genetic mutation in resistant strains to rifampicin and kanamycin.The sensitivity and specificity of whole-genome sequencing for predicting resistance to isoniazid and rifampicin were above 93.44%(95%CI 84.05-98.18), 97.55%(95%CI 96.54-98.33), respectively. The sensitivity and specificity of the whole-genome sequencing technology for predicting resistance to ethambutol and streptomycin was 80.77%(95%CI 60.65-93.45) and 70.18%(95%CI 62.72-70.92), 99.08%(95%CI 98.39-99.52) and 98.18%(95%CI 97.24-98.87), respectively. For the four second-line anti-tuberculosis drugs, the sensitivity of the whole-genome sequencing technology was from 50%(95%CI 6.76-93.24) to 100%(95%CI 88.42-100.00), the specificity was 98.86%(95%CI 98.13-99.36) or above. Conclusion The whole-genome sequencing technology has high application value in the diagnosis of drug-resistant tuberculosis for isoniazid, rifampicin, and ofloxacin, with the higher sensitivity and specificity. But more studies are needed to confirm it's diagnosis value to drug-resistant pulmonary tuberculosis for capreomycin, streptomycinand para-aminosailcylic acid.

Key words: Whole-genome Sequencing, Drug-resistant tuberculosis, Diagnosis, Applied evaluation