PMID- 11352593
OWN - NLM
STAT- MEDLINE
DCOM- 20010816
LR  - 20061115
IS  - 0890-8508 (Print)
IS  - 0890-8508 (Linking)
VI  - 15
IP  - 3
DP  - 2001 Jun
TI  - Specific detection of Stachybotrys chartarum in pure culture using quantitative 
      polymerase chain reaction.
PG  - 129-38
AB  - Research was conducted with laboratory cultures to establish a protocol for the 
      rapid detection and quantitation of the toxigenic fungus Stachybotrys chartarum 
      by means of polymerase chain reaction (PCR). Sequences for the 18 S rRNA gene of 
      S. chartarum were obtained from GenBank and compared against all other available 
      sequences on-line with the Basic Local Alignment Search Tool (BLAST). Two sets of 
      TaqMan primers and one fluorescently labelled probe were designed and tested for 
      selectivity, specificity and sensitivity of detection. A fluorogenic nuclease 
      assay in conjunction with a sequence detector were used for the amplification and 
      quantitation of S. chartarum. The primers designed amplified all S. chartarum 
      isolates tested and did not amplify DNA extracted from other Stachybotrys species 
      or 15 other fungal genera. The primer set selected had a sensitivity of <23 
      template copies. Many S. chartarum samples were initially negative after PCR 
      amplification. Incorporation of an internal positive control in the PCR reaction 
      demonstrated the presence of inhibitors in these samples. PCR inhibitors were 
      removed by dilution or further purification of the DNA samples. The results of 
      this research report on a quantitative PCR (QPCR) method for detection and 
      quantitation of S. chartarum and demonstrate the presence of PCR inhibitors in 
      some S. chartarum isolates.
CI  - Copyright 2001 Academic Press.
FAU - Cruz-Perez, P
AU  - Cruz-Perez P
AD  - Harry Reid Center for Environmental Studies, University of Nevada, Las Vegas, 
      4505 S. Maryland Parkway, Las Vegas, NV 89154-4009, USA.
FAU - Buttner, M P
AU  - Buttner MP
FAU - Stetzenbach, L D
AU  - Stetzenbach LD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - England
TA  - Mol Cell Probes
JT  - Molecular and cellular probes
JID - 8709751
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Ribosomal, 18S)
SB  - IM
MH  - Algorithms
MH  - DNA Primers/metabolism
MH  - Databases, Factual
MH  - Polymerase Chain Reaction/*instrumentation/*methods
MH  - RNA, Ribosomal, 18S/*metabolism
MH  - Software
MH  - Stachybotrys/*metabolism
EDAT- 2001/05/16 10:00
MHDA- 2001/08/17 10:01
CRDT- 2001/05/16 10:00
PHST- 2001/05/16 10:00 [pubmed]
PHST- 2001/08/17 10:01 [medline]
PHST- 2001/05/16 10:00 [entrez]
AID - S0890-8508(01)90347-0 [pii]
AID - 10.1006/mcpr.2001.0347 [doi]
PST - ppublish
SO  - Mol Cell Probes. 2001 Jun;15(3):129-38. doi: 10.1006/mcpr.2001.0347.