Multiplex PCR for the discrimination of A. fumigatus, A. flavus, A. niger and A. terreus

Logotheti, Maria; Kotsovili-Tseleni, A.; Arsenis, G.; Legakis, N.I.

doi:10.1016/j.mimet.2008.10.006

Cited by 33 publications

(22 citation statements)

References 15 publications

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“…Species specific PCR assays were carried out by using reported primers PEPO1, PEPO2 and AnF, AnR were used for specific detection of A. flavus and A. niger respectively [28,29]. Primer sequence and references were given in ( A standard protocol was used to perform nor-1 gene PCR.…”

Section: Species Specific Pcr Assays: Primers and Polymerase Chain Rementioning

confidence: 99%

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India

Priyanka¹,

Ramana²,

Balakrishna³

et al. 2012

AiM

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In the present study, a novel non radioactive digoxigenen labelled PCR-DNA probe was developed targeting nor-1 gene to assess the contamination of aflatoxigenic Aspergillus species in food grain samples from Southern parts of India. The sensitivity of developed PCR-DNA probe was determined to be 10 pg of genomic DNA and 1 pg of purified PCR product. The specificity of the DNA probe was validated by testing against an array of Aspergillus, Fusarium and Penicillium spp. A total of 89 Aspergillus isolates were recovered from 152 grain samples of maize, paddy, and groundnut. Among them, maize had the highest (90%) incidence of toxigenic Aspergillus species. When developed PCR-DNA probe was evaluated onto pure cultures of toxigenic and nontoxigenic Aspergillus species, 60 samples were positive for the nor-1 gene probe. DNA probe results unequivocally matched with the HPLC analysis. In conclusion, the novel PCR-DNA probe developed in this study may find its application in rapid detection of aflatoxin-producing Aspergillus isolates from contaminated cereal grains.

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Section: Species Specific Pcr Assays: Primers and Polymerase Chain Rementioning

confidence: 99%

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India

Priyanka¹,

Ramana²,

Balakrishna³

et al. 2012

AiM

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“…Mitochondrial and rRNA gene regions are the preferred DNA targets for identification of Aspergillus to section level (14,15,24,36). Other approaches tailored to identify Aspergillus to section level are more complex and expensive than our probe-HRM assay, as they require amplification of several targets and inclusion of multiple DNA probes (20,32,44). Two commercial systems make it possible to detect Aspergillus DNA in clinical samples.…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection and Identification of Aspergillus from Lower Respiratory Tract Specimens by Use of a Combined Probe–High-Resolution Melting Analysis

et al. 2012

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Diagnosis of invasive aspergillosis (IA) requires increasingly rapid molecular methods that enable sensitive detection and discrimination between species. We designed and evaluated a real-time PCR-based method that combined melting temperature ( T m ) calling analysis of a specific probe with high-resolution melting analysis of the full amplicon. The test correctly identified 78 isolates of Aspergillus section Fumigati and non- Fumigati sections of Aspergillus with a limit of detection of 10 2 conidia/ml (10 2 fg/ml). No cross-reactivity with other fungi was found. The assay was further validated on lower respiratory tract specimens containing Aspergillus or not. It successfully identified Aspergillus to section level in 56 of 59 specimens. With culture as the gold standard, our assay shows 100% sensitivity and specificity and constitutes an efficient alternative for identification of Aspergillus in lower respiratory tract samples.

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“…Aspergillus flavus diagnosis aims to amplify the target area (1st exon). Logotheti et al (2009) discovered that only 30 isolates of the A. flavus have shown polymerization in size 200 base pairs while ten remaining isolates did not show that bands (Fig. 2).…”

Section: Determination Of Culture Characters For Aspergillus Flavusmentioning

confidence: 97%

Molecularly Diagnostic of Aflatoxigenic Aspergillus flavus Isolated from Nuts

Hamed¹,

Murad²,

Abdul-Rahi³

2016

Research J. of Environmental Toxicology

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Aflatoxigenic fungi like Aspergillus spp., commonly contaminants food, which has different methods to detect their presence. The early detection of food contaminants is quite significant to prevent the hazard on the human health and economic. This study aims to detect the isolation and diagnosis of the companying fungi from some nuts that are available in local markets of Babylon province (Iraq) and explore their ability to produce aflatoxins. The Aspergillus spp., was the most common, followed by each of the Cladosporium spp. and Penicillium spp. The results from isolation and diagnosis of this study showed that Aspergillus flavus was the most visible fungus in all kinds of examined nuts. The highest rate of appearance for this fungus was in pistachios, reaching 70%, whereas the rest of kinds of nuts ranged between 60-10%. The Polymerase Chain Reaction (PCR) technique was utilized for the diagnosis of A. flavus by using a special primer. In this study, the PCR technique was used for the detection of Aflatoxigenic A. flavus and ammonia chemical detection was used to compare this technique.

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Multiplex PCR for the discrimination of A. fumigatus, A. flavus, A. niger and A. terreus

Cited by 33 publications

References 15 publications

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India

Rapid Detection and Identification of Aspergillus from Lower Respiratory Tract Specimens by Use of a Combined Probe–High-Resolution Melting Analysis

Molecularly Diagnostic of Aflatoxigenic Aspergillus flavus Isolated from Nuts

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Multiplex PCR for the discrimination of A. fumigatus, A. flavus, A. niger and A. terreus

Cited by 33 publications

References 15 publications

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing &lt;i&gt;Aspergillus&lt;/i&gt; Species from Major Food Crops Grown in India

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing &lt;i&gt;Aspergillus&lt;/i&gt; Species from Major Food Crops Grown in India

Rapid Detection and Identification of Aspergillus from Lower Respiratory Tract Specimens by Use of a Combined Probe–High-Resolution Melting Analysis

Molecularly Diagnostic of Aflatoxigenic Aspergillus flavus Isolated from Nuts

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A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India

A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India