PMID- 30048818
OWN - NLM
STAT- MEDLINE
DCOM- 20190107
LR  - 20190107
IS  - 1878-3511 (Electronic)
IS  - 1201-9712 (Linking)
VI  - 75
DP  - 2018 Oct
TI  - Rapid method for detecting and differentiating Mycobacterium tuberculosis complex 
      and non-tuberculous mycobacteria in sputum by fluorescence in situ hybridization 
      with DNA probes.
PG  - 1-7
LID - S1201-9712(18)34469-2 [pii]
LID - 10.1016/j.ijid.2018.07.011 [doi]
AB  - OBJECTIVE: In resource-limited tuberculosis-endemic countries, Mycobacterium 
      tuberculosis in sputum is mainly detected by acid-fast bacillus (AFB) staining 
      and the identification of sputum-derived cultures. PCR techniques are practical 
      only in well-resourced laboratories. This study investigated the application of a 
      rapid, simple, and inexpensive fluorescence in situ hybridization (FISH) assay to 
      identify and differentiate M. tuberculosis complex (MTBC) from non-tuberculous 
      mycobacteria (NTM) in sputum. METHODS: The Mycobacterium/Nocardia Genus (MN 
      Genus)-MTBC FISH assay performed in this study utilizes two different DNA probes 
      labeled with different fluorescent molecules that hybridize respectively with 16S 
      rRNA of the genus Mycobacterium and 23S rRNA of MTBC. The assay was tested on 202 
      patient sputum samples in Mangaluru, Karnataka State, India. Sputa were first 
      liquefied and bacteria concentrated before performing the FISH assay and parallel 
      culturing and AFB staining. The identities of cultured bacteria from DNA 
      sequencing were compared with FISH assay findings from corresponding sputa. 
      RESULTS: Of the 202 sputum samples tested, 67 reacted with both MN Genus-specific 
      and MTBC-specific probes, none reacted only with the MTBC-specific probe, and 22 
      reacted only with the MN Genus-specific probe. The FISH assay yielded results in 
      2h and had a limit of detection of 2.2x10(4)CFU/ml in sputum spiked with cultured 
      M. tuberculosis. The diagnostic sensitivity, specificity, and positive and 
      negative predictive values of the FISH assay for MTBC in patient sputa were 
      89.7%, 95.5%, 88.0%, and 92.6%, respectively. NTM were a significant cause of 
      tuberculosis-like infections in Mangaluru. CONCLUSIONS: The MN Genus-MTBC dual 
      probe fluorescence FISH assay previously applied to cultures can also be utilized 
      in resource-limited tuberculosis-endemic countries for rapidly identifying and 
      differentiating MTBC and NTM in sputum samples.
CI  - Copyright (c) 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.
FAU - Baliga, Shrikala
AU  - Baliga S
AD  - Kasturba Medical College, Manipal Academy of Higher Education, Mangaluru, 
      Karnataka, India.
FAU - Murphy, Christina
AU  - Murphy C
AD  - ID-FISH Technology, Palo Alto, CA, USA.
FAU - Sharon, Leesha
AU  - Sharon L
AD  - Kasturba Medical College, Manipal Academy of Higher Education, Mangaluru, 
      Karnataka, India.
FAU - Shenoy, Suchitra
AU  - Shenoy S
AD  - Kasturba Medical College, Manipal Academy of Higher Education, Mangaluru, 
      Karnataka, India.
FAU - Biranthabail, Dhanashree
AU  - Biranthabail D
AD  - Kasturba Medical College, Manipal Academy of Higher Education, Mangaluru, 
      Karnataka, India.
FAU - Weltman, Helena
AU  - Weltman H
AD  - ID-FISH Technology, Palo Alto, CA, USA.
FAU - Miller, Steve
AU  - Miller S
AD  - University of California, San Francisco, CA, USA.
FAU - Ramasamy, Ranjan
AU  - Ramasamy R
AD  - ID-FISH Technology, Palo Alto, CA, USA.
FAU - Shah, Jyotsna
AU  - Shah J
AD  - ID-FISH Technology, Palo Alto, CA, USA. Electronic address: jyotsna@aol.com.
LA  - eng
PT  - Journal Article
DEP - 20180724
PL  - Canada
TA  - Int J Infect Dis
JT  - International journal of infectious diseases : IJID : official publication of the 
      International Society for Infectious Diseases
JID - 9610933
RN  - 0 (DNA Probes)
SB  - IM
MH  - *DNA Probes
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mycobacterium tuberculosis/genetics/*isolation & purification
MH  - Nontuberculous Mycobacteria/genetics/*isolation & purification
MH  - Sputum/*microbiology
MH  - Tuberculosis/*diagnosis
OTO - NOTNLM
OT  - Fluorescence in situ hybridization
OT  - LED fluorescence microscopy
OT  - Mycobacterium tuberculosis
OT  - Non-tuberculous mycobacteria
OT  - Sputum assay
OT  - Tuberculosis diagnosis
EDAT- 2018/07/27 06:00
MHDA- 2019/01/08 06:00
CRDT- 2018/07/27 06:00
PHST- 2018/03/18 00:00 [received]
PHST- 2018/07/07 00:00 [revised]
PHST- 2018/07/09 00:00 [accepted]
PHST- 2018/07/27 06:00 [pubmed]
PHST- 2019/01/08 06:00 [medline]
PHST- 2018/07/27 06:00 [entrez]
AID - S1201-9712(18)34469-2 [pii]
AID - 10.1016/j.ijid.2018.07.011 [doi]
PST - ppublish
SO  - Int J Infect Dis. 2018 Oct;75:1-7. doi: 10.1016/j.ijid.2018.07.011. Epub 2018 Jul 
      24.