Berkala Penelitian Hayati
Vol 19 No 2 (2014): June 2014

PCR APPROACH FOR RAPID DETECTION OF Escherichia coli IN TEMPE USING A SPECIFIC PRIMER

Siti Harnina Bintari (Department of Biology Semarang State University Indonesia, Sekaran, Central Java)
Fidia Fibriana (Department of Biology Semarang State University Indonesia, Sekaran, Central Java)
Dewi Mustikaningtyas (Department of Biology Semarang State University Indonesia, Sekaran, Central Java)
Retno Sri Iswari (Department of Biology Semarang State University Indonesia, Sekaran, Central Java)



Article Info

Publish Date
09 Dec 2014

Abstract

Tempe known as a traditional fermented food originated from Indonesia. It has a unique flavour and texture. It also contains high protein and usually serves to substitute meat, fish, or egg as a complement to rice. The manufacture process of Tempe is quite complex and mostly, the traditional process has not employed the hygienic standard. In the process of Tempe making, there are two critical stages of the whole process; i.e. soaking of soybeans and solid state fermentation by Rhizopus sp. During the process, foodborne pathogen bacteria such as Escherichia coli could contaminate the product of Tempe. The bacterial contamination could be revealed through culture dependent methods which is costly, laborious, and time consuming. Therefore, the culture-independent method such as polymerase chain reaction using a specific primer could be applied to detect target microorganism to save time and labour. In this study, thirty-one Tempe samples collected from different manufacturers in Semarang, Central Java, Indonesia were analysed by PCR. In order to obtain the bacterial genomic DNA, a modified Chelex 100-Microwave method was employed. The results of DNA extraction showed that the method was an applicable method. It gave high quantity and quality of DNA; therefore, it could be applied in the PCR reaction. The DNA samples were employed in PCR for detection of Escherichia coli using Ecoli706F/R. It was found that 27 out of 31 samples were detected having Escherichia coli contamination showed by the presence of the amplified product size 706 bp. The application of this method could significantly reduce costs and time of analysis in the laboratory. Further response after E. coli were detected could be employed, including investigation of the critical factors in Tempe manufacturing process which allowed E. coli contamination.

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Journal Info

Abbrev

jurnal

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology

Description

Berkala Penelitian Hayati is a half yearly international peer reviewed, an open access life science journal. The journal was published by The East Java Biological Society and formerly used the Indonesian language. The first edition of this journal is Vol 1 No 1 in June 1995. It was accredited by ...