A simple polymerase chain reaction-sequencing analysis...fungal

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_Mycopathologia._ (javascript:AL_get(this, 'jour', 'Mycopathologia.')

2006 Oct;162(4):265-71. _Related Articles,_

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A simple polymerase chain reaction-sequencing analysis capable of

identifying multiple medically relevant filamentous fungal species.

_Dean TR_

(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed & cmd=Search & itool=pubmed\

_Abstract & term= " Dean+TR " [Author]) , _Kohan M_

(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed & cmd=Search & itool=pubmed\

_Abstract & term= "

Kohan+M " [Author]) , _Betancourt D_

(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed & cmd=Search & itool=pubmed\

_Abstract & term= " Betancourt+D " [Author])

, _Menetrez MY_

(http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed & cmd=Search & itool=pubmed\

_Abstract & term= " Menetrez+MY " [Author]) .

National Risk Management Research Laboratory, U.S. Environmental Protection

Agency, 109 T.W. Drive, Research Triangle Park, NC, 27711, USA,

dean.timothy@....

Due to the accumulating evidence that suggests that numerous unhealthy

conditions in the indoor environment are the result of abnormal growth of the

filamentous fungi (mold) in and on building surfaces it is necessary to

accurately determine the organisms responsible for these maladies and to

identify them

in an accurate and timely manner. Historically, identification of

filamentous fungal (mold) species has been based on morphological

characteristics, both

macroscopic and microscopic. These methods may often be time consuming and

inaccurate, necessitating the development of identification protocols that are

rapid, sensitive, and precise. To this end, we have devised a simple PAN-PCR

approach which when coupled to cloning and sequencing of the clones allows

for the unambiguous identification of multiple fungal organisms. Universal

primers are used to amplify ribosomal DNA sequences which are then cloned and

transformed into Escherichia coli. Individual clones are then sequenced and

individual sequences analyzed and organisms identified. Using this method we

were capable of identifying Stachybotrys chartarum, Penicillium purpurogenum,

Aspergillus sydowii, and Cladosporium cladosporioides from a mixed culture.

This method was found to be rapid, highly specific, easy to perform, and cost

effective.

PMID: 17039272 [PubMed - in process]