Bacterial Strain and Species Identification

The WAVE^® System has also been used for Species and Strain Identification. Two different applications are described in the following sections.

TB Typing using MIRU-VNTR

Evans and Hawkey et al.¹ have been the first to report the successful use of non–denaturing HPLC on the WAVE System for screening for variations in the number of MIRU-VNTRs in mycobacterial DNA. They conclude:

"Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing of Mycobacterium tuberculosis complex isolates is portable, 100% reproducible, and highly discriminatory. Nondenaturing high-performance liquid chromatography (non-dHPLC) with use of a WAVE microbial analysis system is a promising method of PCR amplicon analysis as it is low cost and requires no preanalysis processing...
Non-dHPLC analysis was demonstrated to be a rapid, low-labor input method for the detection and analysis of MIRU-VNTR amplicons. The combination with non-dHPLC further enhances the utility of MIRU-VNTR typing."

Identification of B. anthracis using DHPLC on the WAVE System

Hurtle et al.² used the WAVE System successfully for the identification of B. anthracis.

"To evaluate DHPLC as a method of identifying B. anthracis, we tested the 16S-23S-specific and gyrA-specific primers, using the B. anthracis reference strains BA1036 and BA1016 with two blind panels of 45 organisms each. Of the 24 B. anthracis strains in the panel tested with the 16S-23S ISR-specific primer set, all were correctly identified. Of the 16 B. anthracis strains in the panel tested with both gyrA-specific primer sets, all strains were correctly identified. Given the high degree of specificity, these results indicate that DHPLC can be used to screen large numbers of samples for B. anthracis.
In summary, we tested a total of 73 members of the B. cereus group to determine if DHPLC could be used to differentiate B. anthracis from B. cereus and B. thuringiensis by analyzing the 16S-23S ISR and a portion of the gyrA gene. In addition, blind panels of 45 samples were used to investigate the capability of DHPLC to identify B. anthracis. We were successful in identifying all the B. anthracis strains in each panel."

References

Evans JT, Hawkey PM, Smith EG, Boese KA, Warren RE and Hong G, Automated High-Throughput Mycobacterial Interspersed Repetitive Unit Typing of Mycobacterium tuberculosis Strains by a Combination of PCR and Nondenaturing High-Performance Liquid Chromatography. J. Clin. Microbiol. 2004, 42(9), 4175-4180 Abstract Library Entry
Hurtle W, Bode E, Kaplan RS, Garrison J, Kearney B, Shoemaker D, Henchal E, Norwood D., Use of denaturing high-performance liquid chromatography to identify Bacillus anthracis by analysis of the 16S-23S rRNA interspacer region and gyrA gene, J. Clin. Microbiol. 2003, 41, 4758-4766. Abstract Library Entry