PMID- 30068538
OWN - NLM
STAT- MEDLINE
DCOM- 20190918
LR  - 20190918
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Print)
IS  - 0095-1137 (Linking)
VI  - 56
IP  - 10
DP  - 2018 Oct
TI  - FISHing Mycobacterium tuberculosis Complex by Use of a rpoB DNA Probe Bait.
LID - 10.1128/JCM.00568-18 [doi]
LID - e00568-18
AB  - Routine staining of sputum specimens does not identify acid-fast bacilli as 
      Mycobacterium tuberculosis with utmost precision, limiting its usability as a 
      confirmatory test for pulmonary tuberculosis. We have combined Ziehl-Neelsen 
      staining and fluorescence in situ hybridization (FISH) to detect M. tuberculosis 
      in sputum specimens. We have developed a new fluorescent oligonucleotide rpoBMTC 
      probe (5'-Alexa-555-AGCGGGGTGATGTCAACCCAG-3') targeting the M. tuberculosis 
      complex rpoB gene. In silico alignment yielded 100% match for M. tuberculosis 
      complex mycobacteria, 66.6% to 47.6% for other bacteria, and no significant hits 
      for viruses and eukaryotes. Negative binding of rpoBMTC probe to the top six 
      respiratory tract bacterial pathogens and to Mycobacterium abscessus and 
      Mycobacterium avium experimentally confirmed its specificity. As for sensitivity, 
      rpoBMTC-FISH detected 10(3) CFU/ml M. tuberculosis as confirmed by successful 
      detection of M. tuberculosis in artificially seeded sputum samples. The 
      application of rpoBMTC-FISH to 116 routine sputum specimens yielded a detection 
      of M. tuberculosis in all of the 31 Ziehl-Neelsen-positive and culture-positive 
      specimens, and no detection of M. tuberculosis in the 85 M. tuberculosis-negative 
      specimens. These data established the proof of concept that rpoBMTC-FISH alone or 
      combined with Ziehl-Neelsen staining can specifically "FISH out" M. tuberculosis 
      complex mycobacteria in sputum samples collected from patients suspected of 
      pulmonary mycobacteriosis. We are implementing this probe for the routine and 
      specific detection of M. tuberculosis complex bacteria in sputum exhibiting 
      acid-fast mycobacteria.
CI  - Copyright (c) 2018 American Society for Microbiology.
FAU - Loukil, Ahmed
AU  - Loukil A
AD  - Aix-Marseille Universite, IRD, MEPHI, IHU Mediterranee-Infection, Marseille, 
      France.
FAU - Kirtania, Prithwiraj
AU  - Kirtania P
AD  - Aix-Marseille Universite, IRD, MEPHI, IHU Mediterranee-Infection, Marseille, 
      France.
FAU - Bedotto, Marielle
AU  - Bedotto M
AD  - Aix-Marseille Universite, IRD, MEPHI, IHU Mediterranee-Infection, Marseille, 
      France.
FAU - Drancourt, Michel
AU  - Drancourt M
AD  - Aix-Marseille Universite, IRD, MEPHI, IHU Mediterranee-Infection, Marseille, 
      France michel.drancourt@univ-amu.fr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180925
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (Bacterial Proteins)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (rpoB protein, Mycobacterium tuberculosis)
RN  - EC 2.7.7.6 (DNA-Directed RNA Polymerases)
SB  - IM
MH  - Bacterial Proteins/*genetics
MH  - Bacteriological Techniques/*methods
MH  - DNA-Directed RNA Polymerases/*genetics
MH  - Fluorescent Dyes
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Microscopy, Fluorescence
MH  - Mycobacterium tuberculosis/genetics/*isolation & purification
MH  - Sensitivity and Specificity
MH  - Sputum
MH  - Tuberculosis, Pulmonary/*diagnosis
PMC - PMC6156301
OTO - NOTNLM
OT  - Mycobacterium tuberculosis
OT  - fluorescence in situ hybridization
OT  - oligonucleotide probe
OT  - rpoB
OT  - tuberculosis
EDAT- 2018/08/03 06:00
MHDA- 2019/09/19 06:00
PMCR- 2019/03/25
CRDT- 2018/08/03 06:00
PHST- 2018/04/05 00:00 [received]
PHST- 2018/07/28 00:00 [accepted]
PHST- 2018/08/03 06:00 [pubmed]
PHST- 2019/09/19 06:00 [medline]
PHST- 2018/08/03 06:00 [entrez]
PHST- 2019/03/25 00:00 [pmc-release]
AID - JCM.00568-18 [pii]
AID - 00568-18 [pii]
AID - 10.1128/JCM.00568-18 [doi]
PST - epublish
SO  - J Clin Microbiol. 2018 Sep 25;56(10):e00568-18. doi: 10.1128/JCM.00568-18. Print 
      2018 Oct.