PENGEMBANGAN METODE DETEKSI KONTAMINASI BAKTERI PATOGEN Escherichia coli DALAM SUSU DENGAN REAL-TIME POLYMERASE CHAIN REACTION (RTi-PCR)
Detection contaminant bacterial pathogen Escherichia coli in milk with Real-Time Polymerase Chain Reaction (RTi-PCR) have been done. The research aimed to develop detection method of E. coli contamination in milk rapidly. The prosedure of research was begun with development method which was consiste...
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| Main Authors: | , |
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| Format: | Theses and Dissertations NonPeerReviewed |
| Published: |
[Yogyakarta] : Universitas Gadjah Mada
2014
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| Subjects: | |
| Online Access: | https://repository.ugm.ac.id/130077/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=70489 |
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| Institution: | Universitas Gadjah Mada |
| Summary: | Detection contaminant bacterial pathogen Escherichia coli in milk with
Real-Time Polymerase Chain Reaction (RTi-PCR) have been done. The research
aimed to develop detection method of E. coli contamination in milk rapidly.
The prosedure of research was begun with development method which
was consisted of E. coli culture enrichment, DNA isolation, analysis of DNA
concentration, and RTi-PCR analysis that was combined with Melting Curve
Analysis (MCA). Then, the research was continued by validation method which
was consisted of specificity test to compare between positive control and negative
control, precision test or repeatability method in 10 positive control, cut off
detection determining, and the last was method application in commercial milk
sample.
The Results from this research showed that RTi-PCR method used
TEco1553 and TecoI754 primer was right method to detect E. coli contamination
in milk with typical melting peak point at 84,50 °C. Specificity test showed that
this method was specific in detecting E. coli in milk. In precision test, all of
positive control resulted melting peak with melting peak point between 84,00 °C �
84,50 °C. This RTi-PCR method could detected contamination of E. coli in milk
with cut off detection until 1,6 CFU/mL. Five commercial milk samples that were
analysed in application method showed a result that commercial milk samples
which sold in traditional market and stall dairy was positive contaminated E. coli.
Key word: Escherichia coli, milk contamination, Real-Time PCR. |
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