A novel DNA chromatography method to discriminate Mycobacterium abscessus subspecies and macrolide susceptibility
Mitsunori Yoshida,
Sotaro Sano,
Jung-Yien Chien,
Hanako Fukano,
Masato Suzuki,
Takanori Asakura,
Kozo Morimoto,
Yoshiro Murase,
Shigehiko Miyamoto,
Atsuyuki Kurashima,
Naoki Hasegawa,
Po-Ren Hsueh,
Satoshi Mitarai,
Manabu Ato,
Yoshihiko Hoshino
Affiliations
Mitsunori Yoshida
Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan
Sotaro Sano
Medical Solutions Vehicle, Kaneka Co. Ltd., Hyogo, Japan
Jung-Yien Chien
Department of Internal Medicine, Pulmonary and Critical Care Medicine, National Taiwan University Hospital, Taipei, Taiwan; Corresponding authors.
Hanako Fukano
Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan
Masato Suzuki
Antimicrobial Resistance Research Center, National Institute of Infectious Diseases, Tokyo, Japan
Takanori Asakura
Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan; Center for Infectious Diseases and Infection Control, School of Medicine, Keio University, Tokyo, Japan
Kozo Morimoto
Respiratory Disease Center, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Tokyo, Japan
Yoshiro Murase
Department of Mycobacterium Reference and Research, the Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, Japan
Shigehiko Miyamoto
Medical Solutions Vehicle, Kaneka Co. Ltd., Hyogo, Japan
Atsuyuki Kurashima
Respiratory Disease Center, Fukujuji Hospital, Japan Anti-Tuberculosis Association, Tokyo, Japan
Naoki Hasegawa
Center for Infectious Diseases and Infection Control, School of Medicine, Keio University, Tokyo, Japan
Po-Ren Hsueh
Departments of Laboratory Medicine and Internal Medicine, National Taiwan University Hospital, National Taiwan University College of Medicine, Taipei, Taiwan
Satoshi Mitarai
Department of Mycobacterium Reference and Research, the Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, Japan
Manabu Ato
Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan
Yoshihiko Hoshino
Department of Mycobacteriology, Leprosy Research Center, National Institute of Infectious Diseases, Toyko, Japan; Corresponding authors.
Background: The clinical impact of infection with Mycobacterium (M.) abscessus complex (MABC), a group of emerging non-tuberculosis mycobacteria (NTM), is increasing. M. abscessus subsp. abscessus/bolletii frequently shows natural resistance to macrolide antibiotics, whereas M. abscessus subsp. massiliense is generally susceptible. Therefore, rapid and accurate discrimination of macrolide-susceptible MABC subgroups is required for effective clinical decisions about macrolide treatments for MABC infection. We aimed to develop a simple and rapid diagnostic that can identify MABC isolates showing macrolide susceptibility. Methods: Whole genome sequencing (WGS) was performed for 148 clinical or environmental MABC isolates from Japan to identify genetic markers that can discriminate three MABC subspecies and the macrolide-susceptible erm(41) T28C sequevar. Using the identified genetic markers, we established PCR based- or DNA chromatography-based assays. Validation testing was performed using MABC isolates from Taiwan. Finding: We identified unique sequence regions that could be used to differentiate the three subspecies. Our WGS-based phylogenetic analysis indicated that M. abscessus carrying the macrolide-susceptible erm(41) T28C sequevar were tightly clustered, and identified 11 genes that were significantly associated with the lineage for use as genetic markers. To detect these genetic markers and the erm(41) locus, we developed a DNA chromatography method that identified three subspecies, the erm(41) T28C sequevar and intact erm(41) for MABC in a single assay within one hour. The agreement rate between the DNA chromatography-based and WGS-based identification was 99·7%. Interpretation: We developed a novel, rapid and simple DNA chromatography method for identification of MABC macrolide susceptibility with high accuracy. Funding: AMED, JSPS KAKENHI
