Comparative study of two typing methods,hsp65 PRA and ITS sequencing, revealed a possible evolutionary link betweenMycobacterium kansasiitype I and II isolates

Abstract

One hundred and ninety-eight clinical isolates of Mycobacterium kansasii collected between 2003 and 2004 in Japan were genotyped by PCR and restriction enzyme analysis (PRA) and 16S-23S internal transcribed spacer (ITS) sequencing. The results demonstrated that clinical isolates of M. kansasii in Japan are almost exclusively of the type I PRA genotype, as is the case in other countries. Although the results of subtyping using the 16S-23S ITS sequence were generally consistent with subtyping using hsp65 PRA, four strains showed a discrepancy between the two methods. Sequence analysis of the hsp65, gyrB and 16S rRNA genes and the ITS sequence of the four strains suggests that they branched from type II and could be considered an ancestral strain of the type I strain. The newly recognized strains were designated as intermediate type I.

Genomic Insights Into the Mycobacterium kansasii Complex: An Update

“…Having the type I-specific hsp65 gene sequence (sequevar I) and type II-specific ITS sequence (sequevar II), strain no. NLA00100521 represents the so-called intermediate type I (I/II), considered a transitional form between environmental type II and human-adapted type I (Iwamoto and Saito, 2005). Whereas, strains K14 and K19 can be identified as atypical type II (IIb) due to their type II ITS sequence and unique, yet most similar to type II, hsp65 sequences (Iwamoto and Saito, 2005).…”

“…NLA00100521 represents the so-called intermediate type I (I/II), considered a transitional form between environmental type II and human-adapted type I (Iwamoto and Saito, 2005). Whereas, strains K14 and K19 can be identified as atypical type II (IIb) due to their type II ITS sequence and unique, yet most similar to type II, hsp65 sequences (Iwamoto and Saito, 2005). Captivatingly, strain K4 had the same unique hsp65 gene sequence, with all the other sequences being characteristic of type I.…”

“…Moreover, M. kansasii isolates with an intermediate type I (I/II) and atypical type II (IIb) have been reported. Whereas, the former had type I-specific sequence of the hsp65 gene and type IIspecific sequence of the spacer region, the latter displayed type II-specific spacer sequence and a unique hsp65 gene sequence (Iwamoto and Saito, 2005). The separateness of the M. kansasii subspecies was further supported by polymorphisms at other genetic loci, including the RNA polymerase gene (rpoB) and the translational elongation factor Tu (tuf ), successfully applied for the differentiation between the subspecies I-VI (Kim et al, 2001;Bakuła et al, 2016).…”

Variable-number tandem repeat markers for Mycobacterium intracellulare genotyping: comparison to the 16S rRNA gene sequencing

“…Characterization of single-nucleotide polymorphisms (SNPs) has been widely used in bacterial identification and genotyping [27][28][29], as well as SNP analysis [30]. As one of the most conserved genes, these mutable bases play an important role in phylogenetic analysis [31].…”

Molecular typing of Mycobacterium kansasii using pulsed-field gel electrophoresis and a newly designed variable-number tandem repeat analysis

“…The genetic structure of M. kansasii has comprehensively been investigated in two late 1990s studies, which allowed the species to be classified into five distinct subtypes (I-V), based on PCR restriction enzyme analysis (PCR-REA) with the major polymorphic tandem repeat (MPTR) probe, pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP) analysis, and PCR-REA of the hsp65 gene 5 , 6 . Somewhat later, two novel types (VI and VII) along with an intermediate I/II and atypical IIB type have been described 7 , 8 .…”