PCR method for the detection of potential ochratoxin-producing Aspergillus species in coffee beans

Sartori, Daniele; Furlaneto, Márcia Cristina; Martins, Mayra Kassawara; Paula, Maria Rosana Ferreira de; Pizzirani-Kleiner, Aline Aparecida; Taniwaki, Marta Hiromi; Fungaro, Maria Helena Pelegrinelli

doi:10.1016/j.resmic.2005.09.008

Cited by 41 publications

(30 citation statements)

References 18 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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“…Once an RAPD or AFLP marker is sequenced, it can be converted into a robust PCR-based marker. Thus, RAPD and AFLP have been applied successfully for revealing specific marker sequences (Schmidt et al 2003(Schmidt et al , 2004bFungaro et al 2004a;Sartori et al 2006). Such sequences have been used to design species-specific primers that allow the identification and detection of some ochratoxigenic species in food samples.…”

Section: Molecular Markers For the Detection Of Ochratoxigenic Fungimentioning

confidence: 99%

“…No cross-reaction was observed with DNA from coffee beans infected with closely related black aspergilli. Similarly, based on RAPD markers, Sartori et al (2006) developed specific primers to detect A. niger (Table 10.1). The primer pair denoted OPX7 372F / OPX7 372R generated an amplicon of 372 bp in all A. niger stricto sensu isolates, and no amplification product was observed in reactions using DNA from related species.…”

Section: Pcr-detection and Quantification Of Ochratoxigenic Species Wmentioning

confidence: 99%

Molecular Diagnosis of Ochratoxigenic Fungi

Sartori

Taniwaki

Iamanaka

et al. 2010

Molecular Identification of Fungi

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Ochratoxin A (OTA) is one of the most abundant food-contaminating mycotoxins. Its presence in several agricultural commodities has been considered a problem worldwide. This toxin is mainly produced by two genera of fungi: Aspergillus and Penicillium. Ochratoxin A has nephrotoxic, immunosuppressive, and carcinogenic effects; consequently, contamination with OTA presents a major risk for human and animal health. Over the last 5 years, several studies have developed PCR-based assays for identifying and quantifying OTA-producing fungi in food samples. The main objective of these assays is to allow the detection of microorganisms capable of producing OTA, prior to ochratoxin production and accumulation. Several of these attempts will be reviewed and discussed in this chapter.

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“…An example of a typical RAPD pattern is given in Figure 1. Because A. ochraceus (now A. westerdijkiae), A. carbonarius and A. niger are the major species for colonizing Brazilian coffee beans and producing OTA our research group developed a multiplex PCR assay (m-PCR) useful to detect the three target fungi species directly from sample of this commodity (Sartori et al, 2006). The m-PCR is a procedure that allows the simultaneous amplification of more than one target sequence in a single PCR reaction, decreasing the number of reaction to be performed to assess the possible presence of different species in a food sample.…”

Section: Molecular Detection Of Ochratoxigenic Fungimentioning

confidence: 99%

An overview on molecular markers for detection of ochratoxigenic fungi in coffee neans

Fungaro

Sartori

2009

Braz. arch. biol. technol.

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PCR method for the detection of potential ochratoxin-producing Aspergillus species in coffee beans

Cited by 41 publications

References 18 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

Molecular Diagnosis of Ochratoxigenic Fungi

An overview on molecular markers for detection of ochratoxigenic fungi in coffee neans

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PCR method for the detection of potential ochratoxin-producing Aspergillus species in coffee beans

Cited by 41 publications

References 18 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

Molecular Diagnosis of Ochratoxigenic Fungi

An overview on molecular markers for detection of ochratoxigenic fungi in coffee neans

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Product

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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