DEVELOPMENT OF FAST PCR SYSTEM FOR AFLR GENE DETECTION
Pichai Chaichanachaichan11,*, Wannaporn Muangsuwan22,, Pattarawan Ruangsuj32,, Kosum
Chansiri43, Srichan Phornchirasilp54, Montri Yasawong62,#
1
Pharmacology and Biomolecular science, Bangkok
2
Department of Biochemistry, Mahidol University, Bangkok
3
Department of Medicine, Srinakarinwirot University, Bangkok
4
Deparment of Pharmacology, Mahidol University, Bangkok
*e-mail: extra_top@hotmail.com, #e-mail: montri.yas@mahidol.ac.th
Abstract
Aflatoxins are secondary metabolite. The toxins were produced from fungal strains in
the genus Aspergillus. Consuming of agricultural products, which were contaminated with
the aflatoxin is cause of aflatoxicosis-nausea, vomiting, muscle cramp, hepatitis and
hepatocarcinoma. There are many methods to detect aflatoxins either quality or quantity
however, the detection spends much time and requires complicated procedures. In this study,
an indirect method for aflatoxin detection was proposed using aflR gene as a biological
marker. The aflR gene sequences of fungal strains in the genus Aspergillus were collected
from NCBI database. The relationship of the gene among the Aspergillus strains was
described by Bayesian inference tree. Conserved regions of the gene were determined and
then utilized for primers design. The aflR-primers were validated under a standard PCR
protocol. There were four protocols for fast PCR system had developed. Only protocol A was
successfully amplified the aflR amplicon. The fast PCR protocol was faster than the standard
PCR protocol around 2.6 times. The fast PCR system may apply or incorporate with other
techniques which have to enrich DNA target before detection of the results such as DNA
biosensors.
Keywords: aflatoxin, aflR, fast PCR system