[Evaluation PCR-reverse line hybridization assay for detection and identification of Mycobacterium species: a multicenter study].
OBJECTIVE
To evaluate the PCR-reverse line dot hybridization assay (RLB) based on 16S-23S rDNA internal transcribed spacer (ITS) sequence for detection and identification of mycobacterium species.


METHODS
This study was performed in five different centers simultaneously, and the performance of PCR-RLB was estimated and verified by detection of 60 reference strains belonging to 50 mycobacterium species, 10 nonmycobacterial speices and 383 clinical isolates identified by immunochromatographic assay (ICA) combined sequencing and gas chromatography (GC).


RESULTS
The genus-specific probe hybridizied with the amplification of PCR of all mycobacteria and species-specific probes hybridized only with corresponding species, however, neither of them hybridized with the amplification of PCR of nonmycobacterial speices. Compared with ICA combined sequencing and GC, accuracy of PCR-RLB was nearly 100%, and PCR-RLB identified 380 of 383 in species-level successfully, including 11 mixed clinical isolates which could not be identified with other assays correctly.


CONCLUSIONS
The mycobacterial PCR- RLB based on 16S-23S rDNA ITS is rapid, convenient, sensitive and specific, and practical, and it is promising to be applied in clinical diagnosis.
Wang
Ya-Juan
YJ
Center of Infection Disease, West China Hospital, Sichuan University, Chengdu 610041, China.
Xiong
Li-Kuan
LK
Feng
Ping
P
D021903:DNA, Ribosomal Spacer / Q000235:genetics; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009164:Mycobacterium Infections / Q000382:microbiology; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D012680:Sensitivity and Specificity
D021903:DNA, Ribosomal Spacer; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S