Development of a PCR assay for amplification of mating-type loci of Alternaria spp. and related fungi.

2013 ◽  
Vol 65 (1) ◽  
pp. 69-78 ◽  
Author(s):  
Philipp B. Gannibal ◽  
Igor A. Kazartsev
Keyword(s):  
Mating Type ◽  
Pcr Assay

scholarly journals The MAT1 Locus of Histoplasma capsulatum Is Responsive in a Mating Type-Specific Manner

2007 ◽  
Vol 6 (4) ◽  
pp. 616-621 ◽  
Author(s):  
Meggan Bubnick ◽  
A. George Smulian
Keyword(s):  
Mating Type ◽  
Histoplasma Capsulatum ◽  
Genomic Sequence ◽  
Pathogenic Fungus ◽  
Clinical Isolates ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Specific Manner ◽  
Specific Pcr Assay ◽  
Syntenic Analysis

ABSTRACT Recombination events associated with sexual replication in pathogens may generate new strains with altered virulence. Histoplasma capsulatum is a mating-competent, pathogenic fungus with two described phenotypic mating types, + and −. The mating (MAT) locus of H. capsulatum was identified to facilitate molecular studies of mating in this organism. Through syntenic analysis of the H. capsulatum genomic sequence databases, a MAT1-1 idiomorph region was identified in H. capsulatum strains G217B and WU24, and a MAT1-2 idiomorph region was identified in the strain G186AR. A mating type-specific PCR assay was developed, and two clinical isolates of opposite genotypic mating type, UH1 and VA1, were identified. A known − mating type strain, T-3-1 (ATCC 22635), was demonstrated to be of MAT1-2 genotypic mating type. The clinical isolates UH1 and VA1 were found to be mating compatible and also displayed mating-type-dependent regulation of the MAT transcription factors in response to extracts predicted to contain mating pheromones. These studies support a role for the identified MAT1 locus in determining mating type in H. capsulatum.

scholarly journals Genetic Relationships Among Magnaporthe poae Isolates from Turfgrass Hosts and Relative Susceptibility of ‘Penncross’ and ‘Penn A-4’ Creeping Bentgrass

Plant Disease ◽  
2006 ◽  
Vol 90 (12) ◽  
pp. 1531-1538 ◽  
Author(s):  
L. P. Tredway
Keyword(s):  
Mating Type ◽  
Creeping Bentgrass ◽  
Type A ◽  
Kentucky Bluegrass ◽  
Its Sequences ◽  
Growth Chamber ◽  
Poly Merase Chain Reaction ◽  
Pcr Assay ◽  
Annual Bluegrass ◽  
Magnaporthe Poae

Isolates of Magnaporthe poae from turfgrass hosts were analyzed for mating type, genetic relatedness according to ITS sequences, reaction to a previously developed species-specific poly-merase chain reaction (PCR) assay, and virulence on two creeping bentgrass cultivars in growth chamber experiments. Analysis of internal transcribed spacer (ITS) sequences revealed three clades, designated A, B, and C. Clade A contained isolates of both mating types from creeping bentgrass, annual bluegrass, and Kentucky bluegrass. Clade B contained only mating type ‘A’ isolates from annual bluegrass, whereas Clade C contained only mating type ‘a’ isolates from creeping bentgrass. The M. poae PCR assay failed to positively identify several North Carolina isolates from annual bluegrass and creeping bentgrass. M. poae isolates from Kentucky blue-grass were most virulent toward creeping bentgrass in growth chamber experiments. Although isolates of M. poae are not host specific, certain ITS clades may have a limited host or geographical range. The improved creeping bentgrass cv. Penn A-4 was more susceptible to summer patch than cv. Penncross. Additional research is needed to develop methods for accurate diagnosis of summer patch and other patch diseases in creeping bentgrass and to determine how creeping bentgrass cultivars vary in their susceptibility to these root pathogens.

scholarly journals Some evidence for skewed mating type distribution in Iranian populations of Rhynchosporium commune, the cause of barley scald disease

2016 ◽  
Vol 56 (3) ◽  
pp. 237-243 ◽  
Author(s):  
Mahdi Arzanlou ◽  
Kaivan Karimi ◽  
Fariba Mirabi
Keyword(s):  
Mating Type ◽  
Spatial Scales ◽  
Genotypic Diversity ◽  
Sexual Cycle ◽  
Pcr Assay ◽  
Unequal Distribution ◽  
Equal Distribution ◽  
Multiplex Pcr Assay ◽  
The World ◽  
Mating Type Genes

AbstractRhynchosporium commune(formerly known asRhynchosporium secalis), the causal agent of scald disease on barley, is known to spread asexually by splash dispersed conidia. However, there are multiple lines of evidence for the possibility of a clandestine sexual cycle occurrence in this species including extensive genotypic diversity, equal distribution of mating type alleles across the world and expression of mating type genes. In the current study, the potential for the occurrence of a sexual cycle amongst the Iranian population ofR. communewas assessed by analyzing distribution and frequency of the mating type alleles at both micro and macro-spatial scales. A total of 95 single-conidialR. communeisolates were obtained from different barley fields in Kurdistan province. Previously designed primers were applied in a multiplex PCR assay to study distribution and frequency of the mating type alleles within and between populations. Totally, 67 isolates were determined asMAT1-1and the remaining 28 isolates asMAT1-2throughout the sampling counties. The results obtained at a macro-spatial scale revealed that unlike Kamyaran county (bothMAT1-1andMAT1-2at an equal ratio), an unequal distribution of mating type genes was dominant amongR. communeisolates in both Mariwan and Dehgolan counties. Our findings support a predominantly asexual reproduction for Mariwan and Dehgolan counties and the possibility of sexual stage occurrence in Kamyarna county.

A new duplex PCR assay for the rapid screening of mating-type idiomorphs of pathogenic Sporothrix species

2021 ◽  
Author(s):  
Jamile Ambrósio de Carvalho ◽  
Breno Gonçalves Pinheiro ◽  
Ferry Hagen ◽  
Sarah Santos Gonçalves ◽  
Ricardo Negroni ◽  
...  
Keyword(s):  
Mating Type ◽  
Rapid Screening ◽  
Duplex Pcr ◽  
Pcr Assay ◽  
Duplex Pcr Assay

1089: Prognostic Value of Molecular Staging of Bladder Cancer with RT -PCR Assay for KRT20: Results at 5 Year-Follow-up

2007 ◽  
Vol 177 (4S) ◽  
pp. 360-360
Author(s):  
Ana Agud ◽  
Maria J. Ribal ◽  
Lourdes Mengual ◽  
Mercedes Marin-Aguilera ◽  
Laura Izquierdo ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Prognostic Value ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Molecular Staging

A Hemi-nested, Allele Specific, Whole Blood PCR Assay for the Detection of the Factor V Leiden Mutation

1997 ◽  
Vol 77 (06) ◽  
pp. 1154-1155 ◽  
Author(s):  
Gary D Sinclair ◽  
Sandra Low ◽  
Man-Chiu Poon
Keyword(s):  
Whole Blood ◽  
Protein C ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Restriction Enzyme Digestion ◽  
Factor V ◽  
Pcr Assay ◽  
Specific Primers ◽  
Factor V Leiden Mutation ◽  
Allele Specific

SummaryWe describe a novel hemi-nested, allele specific whole blood PCR assay for detection of the factor V Leiden mutation associated with the plasma defect, activated protein C resistance. This assay utilizes 5 μl of whole blood without prior DNA extraction. The hemi-nested design, employing an outer primer pair in combination with nested, allele specific primers obviates the need for restriction enzyme digestion. PCR reactions are analysed directly on agarose or polyacrylamide minigels. The assay confirmed the genotypes of 50 individuals previously categorized by PCR and Mnll digestion, and has been subsequently utilized in the genotyping of 445 individuals referred for thrombosis studies.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

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