Biomedical Engineering Reference
In-Depth Information
made use of the forward primer described in Niessen and Vogel (1998)
which was combined with reverse primers binding specifi cally to the
intron region of the tri5 gene in the respective species. Using
tri
5 gene
sequence Strausbaugh et al. (2005) set up a TaqMan™ quantitative real-time
PCR assay for quantifi cation of
F. culmorum
in wheat roots and in barley
plants.
Stachybotrys chartarum
(syn.
S. atra)
was identifi ed as a producer of
the highly potent macrocyclic trichothecene satratoxin in “sick building
syndrome” households (Etzel et al
.
1998). Strauss and Wong published
the DNA sequence of the
tri5
gene of
Stachybotrys chartarum
(GenBank,
direct submission (1998), accession no.AF053926). The sequence was
subsequently used by Peltola et al. (2002) to design a pair of specifi c PCR
primers for the identifi cation of the toxigenic subgroup 1 of
S. chartarum
.
Non toxic strains and strains with low toxicity were combined in subgroup
2. These fungi did not give a product with the primer pair which thus was
useful to distinguish the two taxonomical groups in
S. chartarum
. Presence
of two lineages in this fungus was also found by Koster et al. (2003), who
described two phylogenetically distinct lineages when testing genotypic
variation of
tri
5, ITS and two housekeeping genes in a geographically
diverse set of isolates. Dean et al
.
(2005) developed another pair of
tri5
-
based primers for identifi cation of
S. chartarum
which were combined with
primers specifi c for
A. versicolor, P. purpurogenum
and
Cladosporium
spp.,
respectively, in a multiplex PCR assay to check buildings for the potential
presence of mycotoxin producing fungi in indoor environments. Besides
tri5
, other genes from the trichothecene biosynthesis cluster were used
to design species- and group specifi c PCR primers. A group specifi c PCR
assay for the detection of trichothecene producing
Fusarium
spp. involving
primers binding to the
tri6
gene (transcription factor), was set up by Bluhm
et al. (2002) and also used the system together with primers for sensitive
detection of fumonisin producers. Bakan et al. (2002) applied intergenic
sequences between the
tri5
and
tri6
genes to distinguish between high
and low DON producing
F. culmorum
isolates. Testing of 17 and 13 isolates
of high and low DON producers, respectively, revealed 100% correlation
between producer type and the size of amplicon produced. In order to
differentiate DON and NIV producing chemotypes in
F. graminearum
,
Lee et al. (2001) designed primers hybridising adjacent to an inserted
region present in the tri7 gene of the fungus. A genus specifi c primer pair
hybridising to sequences within the
tri13
gene was published by Demeke
et al. (2005), who screened 85 samples of wheat, barley, oats and corn
from Canada and found good correlation between PCR detection of
F.
graminearum
biomass and presence of DON in the samples. A multiplex
PCR assay was developed for grouop specifi c detection of Trichothecene
producing fungi was developed by targeting tri5 and tri6 genes along with
Fumonisin producing fungi (Ramana et al. 2011) and developed system
