Reliable identification of mycobacterial species by PCR-restriction enzyme analysis (PRA)-hsp65 in a reference laboratory and elaboration of a sequence-based extended algorithm of PRA-hsp65patterns

Table 4 BstEII and HaeIII fragment lengths (base pairs) for 30 isolates with new patterns by PRA-hsp65.

Species^a	PRA-hsp65	N	Fragment BstEII	Length (bp) HaeIII
*M. arupense*	NP17	5	320-115	145-75-60
M. avium	NP10	1	320-115	140-90-60
M. cosmeticum	NP6	1	320-115	150-95-80
M. fortuitum	NP12	1	235-120-85	140-120-100-55
M. fortuitum	NP19	1	235-120-100	145-140-100-55
*M. gordonae*	NP1	11	235-120-100	130-110-95
M. gordonae	NP3	1	320-130	130-60
M. gordonae	NP13	1	235-120-85	130-90
M. gordonae	NP22	1	235-130-85	160-90-60
M. mageritense	NP5	1	240-130-85	145-100-50
M. nonchromogenicum	NP4	1	235-120-85	145-80-60
*M. sherrisii*	NP14	3	235-120-85	145-130
*M. terrae*	NP11	2	235-210	140-115-70

^a Species identification based on sequencing of hsp65 gene. Bold indicates sequences submitted to GenBank and patterns included in the updated PRA-hsp65 algorithm (see Figures 1, 2 and 3). GenBank accession numbers: NP1, EF601222; NP11, EF601223; NP14, AY365190 [23]; NP17, DQ168662 [18].
All isolates with new PRA-hsp65 profiles were cultured from sputum, with the following exceptions: NP1: urine (2), feces, liver biopsy and unknown (one each); NP17: unknown (2).

ISSN: 1471-2180