Comparison of Nucleotide MALDI-TOF MS with Xpert MTB/RIF for Rifampicin Susceptibility Identification and Associated Risk Factors of Rifampicin Resistance Among Drug Resistant Mycobacterium tuberculosis

Song Song,1,* Honghong Xu,1,* Jiawei Cao,2,* Guanghong Wu,2 Haiyan Sun,1 Xiaoqi Dai,1 Xuekui Li,1 Meng Chen,1 Menghan Zhang,1 Yueming Yan,1 Jingfeng Tong,3 Zhongdong Wang1 1Institute of Tuberculosis Control, Qingdao Preventive Medicine Research Institute, Qingdao Municipal Center for Disease Control and Prevention, Qingdao, People’s Republic of China; 2Department of Laboratory Medicine, Qingdao Chest Hospital, Qingdao, Shandong, People’s Republic of China; 3Department of Medical Affairs, Shanghai Conlight Medical Co., Ltd., Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhongdong Wang, Institute of Tuberculosis Control, Qingdao Preventive Medicine Research Institute, Qingdao Municipal Center for Disease Control and Prevention, No. 8, Fengxian Road, Jinhu Road Street, Shinan District, Qingdao City, Shandong Province, 266000, People’s Republic of China, Email [email protected]: Nucleotide-based matrix-assisted laser desorption ionization time-of-flight mass spectrometry (nucleotide MALDI-TOF MS) is an emerging molecular technology used for the diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB)and its drug resistance. This study aimed to compare the ability of nucleotide MALDI-TOF MS to detect rifampicin (RIF) resistance in drug-resistant TB (DR-TB) patients with Xpert MTB/RIF and to analyze the disparate results individually. Additionally, potential factors associated with rifampicin resistance among DR-TB patients in Qingdao were investigated.Patients and Methods: A retrospective study was conducted at Qingdao Chest Hospital, and patients with DR-TB were enrolled. Corresponding frozen isolates were recovered and subjected to nucleotide MALDI-TOF MS, Xpert MTB/RIF, and phenotypic drug susceptibility testing (pDST). Sanger sequencing was performed for the discordant results of nucleotide MALDI-TOF MS and Xpert MTB/RIF. Univariate and multivariate logistic regression analyses were used to identify potential factors associated with rifampicin resistance among patients with DR-TB.Results: A total of 125 patients with DR-TB (18.8%, 125/668) were enrolled in this study from May 1 to July 31, 2023. Rifampicin-resistant (DR-TB/RR, 29) and rifampicin-sensitive (DR-TB/RS, 96) groups were divided according to the pDST results. Nucleotide MALDI-TOF MS performed better than Xpert MTB/RIF in terms of sensitivity, specificity, accuracy, and agreement with pDST. Only six cases had inconsistent results, and the sequencing results of five cases were identical to nucleotide MALDI-TOF MS. Furthermore, chest pain (aOR=12.84, 95% CI, 2.29– 91.97, p=0.005), isoniazid sensitivity (aOR=0.14, 0.02– 0.59, p=0.013), and ethambutol sensitivity (aOR=0.02, 0.00– 0.10, p=0.000) were potential factors associated with rifampicin resistance among DR-TB patients in Qingdao.Conclusion: The overall concordance between nucleotide MALDI-TOF MS and Xpert MTB/RIF was 95.2%, with the former performing better in determining rifampicin susceptibility among DR-TB cases in Qingdao. Chest pain, isoniazid, and ethambutol resistance might be factors associated with RIF resistance among patients with DR-TB in Qingdao.Keywords: MTB, rifampicin, nucleotide MALDI-TOF MS, Xpert MTB/RIF, comparison, risk factors