Establishment of cross-priming amplification for point-of-care detection of

To simplify sputum sample preparation steps and achieve point-of-care testing (POCT) of Evaluate the liquefying ability of high-concentration guanidine thiocyanate (GTC) for sputum and the effectiveness of MTB inactivation; establish a rapid detection system for MTB and NTM based on CPA technology using EasyNAT integrated detection tubes, with the left amplification zone specific to MTB CPA amplification and the right amplification zone specific to both MTB and NTM CPA amplification. Suspected pulmonary tuberculosis (PTB) patients or patients diagnosed as suspected NTM pulmonary infections specimens collected from the Second Hospital of Longyan, Fujian Province, from September 2022 to September 2023, acid-fast bacilli (AFB) smear, quantitative real-time PCR (RT-PCR) and CPA-POCT were performed. The kappa coefficients was used to evaluate the consistency between the RT-PCR and CPA-POCT. The liquefaction effect of 6M GTC on sputum was equivalent to 4 % NaOH, and no MTB growth was observed in the Lowenstein-Jensen medium of sputum samples treated with 6M GTC incorporating the H37Rv strain. The newly established CPA-POCT method showed good agreement with RT-PCR with a positive compliance rate of 86.27 %, a negative compliance rate of 89.36 %, an overall compliance rate of 87.75 %, and a Kappa coefficients of 0.786 (P < 0.05). 6M GTC can liquefy sputum and render MTB non-viable, eliminating the need for Mycobacterium nucleic acid testing in BSL-2 laboratories; the newly established CPA method can rapidly and accurately distinguish MTB and NTM in the form of POCT, with simple and fast operation, suitable for promotion and application in grassroots medical institutions and remote rural areas.

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Déclaration de conflit d'intérêts

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Wenbin Huang reports financial support was provided by Fujian Society of Laboratory Medicine and the Joint Innovation Project of the National (Fujian) Gene Testing Technology Application Demonstration Center. Yuankui Lin is employed by company “Ustar Biotechnologies (Hangzhou) Ltd.” If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.