PMID- 22088321
OWN - NLM
STAT- MEDLINE
DCOM- 20120416
LR  - 20220716
IS  - 1873-281X (Electronic)
IS  - 1472-9792 (Print)
IS  - 1472-9792 (Linking)
VI  - 91 Suppl 1
DP  - 2011 Dec
TI  - Rapid DNA extraction for specific detection and quantitation of Mycobacterium 
      tuberculosis DNA in sputum specimens using Taqman assays.
PG  - S43-8
LID - 10.1016/j.tube.2011.10.009 [doi]
AB  - Rapid tuberculosis (TB) detection is critical for disease control, and further 
      quantitation of Mycobacterium tuberculosis (Mtb) in sputum is valuable for 
      epidemiological and clinical studies. We evaluated a simple, robust and 
      cost-efficient in-house DNA extraction and downstream Taqman approach for 
      detection and quantitation of Mtb genomes from sputum of newly-diagnosed TB 
      patients and non-TB controls. DNA was extracted using guanidine isothiocyanate 
      and silica-based spin columns in less than 2 h, stored frozen, and Taqman assays 
      were used to detect Mtb with IS6110 and quantify it targeting RD1 and IS1081. The 
      Taqmans had a sensitivity >95% in 108 culture-confirmed TB patients and 
      specificity of 100% in 43 non-TB controls. Genome counts were correlated with the 
      Mycobacterial Growth Indicator Tubes' (MGIT) time-to-detection values (1/TTD x 
      1000; rho = 0.66; p < 0.001) in 91 TB patients (33 excluded with MGIT 
      contamination). This linear relationship was nearly identical between 
      mycobacteria isolated from sputum and H37Rv Mtb grown in-vitro to its log phase. 
      TB treatment between 3 and 7 days was associated with lower 1/TTD x 1000 values 
      but not with genome counts. Together, our protocol provides rapid, specific, 
      inexpensive and quantitative detection of Mtb DNA in fresh or stored sputa making 
      it a robust tool for prompt TB diagnosis, and with potential use for clinical and 
      epidemiologic studies.
CI  - Copyright (c) 2011 Elsevier Ltd. All rights reserved.
FAU - Gomez, Diana I
AU  - Gomez DI
AD  - Division of Epidemiology, School of Public Health in Brownsville, University of 
      Texas Health Science Center at Houston, 80 Fort Brown, SPH Bldg, Brownsville, 
      Texas 78520, USA.
FAU - Mullin, Caroline S
AU  - Mullin CS
FAU - Mora-Guzman, Francisco
AU  - Mora-Guzman F
FAU - Crespo-Solis, J Gonzalo
AU  - Crespo-Solis JG
FAU - Fisher-Hoch, Susan P
AU  - Fisher-Hoch SP
FAU - McCormick, Joseph B
AU  - McCormick JB
FAU - Restrepo, Blanca I
AU  - Restrepo BI
LA  - eng
GR  - R21 AI064297/AI/NIAID NIH HHS/United States
GR  - P20 MD000170/MD/NIMHD NIH HHS/United States
GR  - R21 AI079624/AI/NIAID NIH HHS/United States
GR  - P20 MD000170-04/MD/NIMHD NIH HHS/United States
GR  - R01 AI116039/AI/NIAID NIH HHS/United States
GR  - 1 R21AI064297-01-A1/AI/NIAID NIH HHS/United States
GR  - R21 AI064297-01A1/AI/NIAID NIH HHS/United States
GR  - R21 AI079624-01A1/AI/NIAID NIH HHS/United States
PT  - Evaluation Study
PT  - Journal Article
PT  - Multicenter Study
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20111115
PL  - Scotland
TA  - Tuberculosis (Edinb)
JT  - Tuberculosis (Edinburgh, Scotland)
JID - 100971555
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - Adult
MH  - Case-Control Studies
MH  - DNA, Bacterial/*isolation & purification
MH  - Female
MH  - Genome, Bacterial
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Mycobacterium tuberculosis/genetics/*isolation & purification
MH  - Real-Time Polymerase Chain Reaction/methods
MH  - Sensitivity and Specificity
MH  - Specimen Handling/methods
MH  - Sputum/*microbiology
MH  - Time Factors
MH  - Tuberculosis, Pulmonary/*diagnosis
PMC - PMC3248966
MID - NIHMS338900
OTO - NOTNLM
OT  - PCR inhibitors
OT  - diagnosis
OT  - quantitation
OT  - real-time quantitative PCR
OT  - sputum
OT  - tuberculosis
COIS- Competing interest: None declared.
EDAT- 2011/11/18 06:00
MHDA- 2012/04/17 06:00
PMCR- 2012/12/01
CRDT- 2011/11/18 06:00
PHST- 2011/11/18 06:00 [entrez]
PHST- 2011/11/18 06:00 [pubmed]
PHST- 2012/04/17 06:00 [medline]
PHST- 2012/12/01 00:00 [pmc-release]
AID - S1472-9792(11)00179-X [pii]
AID - 10.1016/j.tube.2011.10.009 [doi]
PST - ppublish
SO  - Tuberculosis (Edinb). 2011 Dec;91 Suppl 1:S43-8. doi: 10.1016/j.tube.2011.10.009. 
      Epub 2011 Nov 15.