A rapid and reliable method for the specific detection of aflatoxigenic fungi in groundnut and rice samples

Aflatoxin contamination is a serious food safety and economic concern for the agriculture and processing industry throughout the world. Aflatoxins are the most potent carcinogens; therefore, an early detection of aflatoxin‐producing fungi is essential for ensuring food safety. In the present work, a rapid and sensitive PCR method for the detection of potential aflatoxin‐producing molds in groundnut and rice samples was developed. This PCR assay can detect the aflatoxigenic fungi in the contaminated samples directly without the need of fungal enrichment. The results showed a specific PCR amplification product in aflatoxigenic fungi (Aspergillus parasiticus and Aspergillus flavus) contaminated groundnut and rice samples even at a low spore count. In contrast, no amplification products were observed in nonaflatoxigenic fungi (Aspergillus oryzae) contaminated samples as well as in healthy groundnut and rice samples. Quantification of aflatoxins for the validation corelated well with the amplified DNA bands for aflatoxigenic fungal samples. PRACTICAL APPLICATIONS: The present paper describes a rapid and reliable PCR method for the detection of potential aflatoxin‐producing molds in groundnut and rice samples. The developed method is sensitive and rapid, as it detects the aflatoxigenic fungi in the contaminated samples directly without the need of fungal enrichment. The developed method is sensitive because this process detects the fungal infestation at a concentration as low as 10² spores/g. This method can be used to detect the presence of aflatoxigenic fungi specifically, even before the production of aflatoxin to ensure food safety.

[1]  H. Oberoi,et al.  Validation of PCR based detection system for aflatoxin producing molds. , 2016, Indian journal of experimental biology.

[2]  Anong Bintvihok,et al.  A Rapid and Sensitive Detection of Aflatoxin-producing Fungus Using an Optimized Polymerase Chain Reaction (PCR) , 2016, Toxicological research.

[3]  S. Samanta,et al.  Occurrence of Aflatoxigenic A. flavus in Stored Rice and Rice Based Products of Coastal Odisha, India , 2014 .

[4]  M. Skrinjar,et al.  Identification of aflatoxigenic fungi using polymerase chain reaction-based assay , 2014 .

[5]  V. Sureshkumar,et al.  Survey of market samples of food grains and grain flour for Aflatoxin B1 contamination , 2013 .

[6]  R. Levin.  PCR detection of aflatoxin producing fungi and its limitations. , 2012, International journal of food microbiology.

[7]  C. Vázquez,et al.  Specific detection and quantification of Aspergillus flavus and Aspergillus parasiticus in wheat flour by SYBR® Green quantitative PCR. , 2011, International journal of food microbiology.

[8]  K. Reddy,et al.  Detection of Aspergillus spp. and aflatoxin B1 in rice in India. , 2009, Food microbiology.

[9]  C. Vázquez,et al.  Highly sensitive PCR-based detection method specific for Aspergillus flavus in wheat flour , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[10]  L. Rocha,et al.  DISTRIBUTION OF FUNGI AND AFLATOXINS IN A STORED PEANUT VARIETY , 2008 .

[11]  J. Vytřasová,et al.  Detection of aflatoxigenic fungi in feeds using the PCR method , 2008, Folia Microbiologica.

[12]  S. Shahsavandi,et al.  Application of PCR on detection of aflatoxinogenic fungi , 2007 .

[13]  A. Chandrashekar,et al.  PCR-restriction fragment length analysis of aflR gene for differentiation and detection of Aspergillus flavus and Aspergillus parasiticus in maize. , 2004, International journal of food microbiology.

[14]  John E. Linz,et al.  Clustered Pathway Genes in Aflatoxin Biosynthesis , 2004, Applied and Environmental Microbiology.

[15]  K. Bhattacharya,et al.  Deteriorative changes of maize, groundnut and soybean seeds by fungi in storage , 2004, Mycopathologia.

[16]  R. Geisen,et al.  Monitoring the Production of Aflatoxin B1 in Wheat by Measuring the Concentration of nor-1 mRNA , 2003, Applied and Environmental Microbiology.

[17]  A. Dobson,et al.  PCR-based detection and quantification of mycotoxigenic fungi , 2002 .

[18]  R. Chiou,et al.  Polymerase chain reaction-mediated characterization of molds belonging to the Aspergillus flavus group and detection of Aspergillus parasiticus in peanut kernels by a multiplex polymerase chain reaction. , 2002, Journal of food protection.

[19]  F. X. Bosch,et al.  Aflatoxin, hepatitis and worldwide liver cancer risks. , 2002, Advances in experimental medicine and biology.

[20]  G. Criseo,et al.  Differentiation of aflatoxin‐producing and non‐producing strains of Aspergillus flavus group , 2001, Letters in applied microbiology.

[21]  Y. Kumeda,et al.  Heteroduplex Panel Analysis, a Novel Method for Genetic Identification of Aspergillus SectionFlavi Strains , 2001, Applied and Environmental Microbiology.

[22]  J. Pitt,et al.  Toxigenic fungi: which are important? , 2000, Medical mycology.

[23]  A. Dobson,et al.  The use of reverse transcription-polymerase chain reaction (RT-PCR) for monitoring aflatoxin production in Aspergillus parasiticus 439. , 2000, International journal of food microbiology.

[24]  Grimm,et al.  A PCR‐ELISA for the detection of potential fumonisin producing Fusarium species , 1998, Letters in applied microbiology.

[25]  W. Holzapfel,et al.  Detection of aflatoxinogenic fungi in figs by a PCR reaction. , 1997, International journal of food microbiology.

[26]  N. Paster,et al.  Detection of aflatoxigenic molds in grains by PCR , 1996, Applied and environmental microbiology.

[27]  Glasel Ja.  Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. , 1995 .

[28]  J. Glasel.  Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. , 1995, BioTechniques.

[29]  H. Sandermann,et al.  A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. , 1992, Nucleic acids research.