title: Computational approach involving use of the internal transcribed spacer 1 region for identification of Mycobacterium species.
abstract: The rapid and reliable identification of clinically significant Mycobacterium species is a challenge for diagnostic laboratories. This study evaluates a unique sequence-dependent identification algorithm called MycoAlign for the differential identification of Mycobacterium species. The MycoAlign system uses pan-Mycobacterium-specific primer amplification in combination with a customized database and algorithm. The results of testing were compared with conventional phenotypic assays and GenBank sequence comparisons using the 16S rRNA target. Discrepant results were retested and evaluated using a third independent database. The custom database was generated using the hypervariable sequences of the internal transcribed spacer 1 (ITS-1) region of the rRNA gene complex from characterized Mycobacterium species. An automated sequence-validation process was used to control quality and specificity of evaluated sequence. A total of 181 Mycobacterium strains (22 reference strains and 159 phenotypically identified clinical isolates) and seven nonmycobacterial clinical isolates were evaluated in a comparative study to validate the accuracy of the MycoAlign algorithm. MycoAlign correctly identified all referenced strains and matched species in 94% of the phenotypically identified Mycobacterium clinical isolates. The ITS-1 sequence target showed a higher degree of specificity in terms of Mycobacterium identification than the 16S rRNA sequence by use of GenBank BLAST. This study showed the MycoAlign algorithm to be a reliable and rapid approach for the identification of Mycobacterium species and confirmed the superiority of the ITS-1 region sequence over the 16S rRNA gene sequence as a target for sequence-based species identification.
authors.affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198-6495, USA.
authors.forename: Amr M
authors.initials: AM
authors.lastname: Mohamed
authors.forename: Dan J
authors.initials: DJ
authors.lastname: Kuyper
authors.forename: Peter C
authors.initials: PC
authors.lastname: Iwen
authors.forename: Hesham H
authors.initials: HH
authors.lastname: Ali
authors.forename: Dhundy R
authors.initials: DR
authors.lastname: Bastola
authors.forename: Steven H
authors.initials: SH
authors.lastname: Hinrichs
mesh_terms: D001483:Base Sequence; D021903:DNA, Ribosomal Spacer; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics
chemical_list: D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S