Molecular Characterization of Listeria monocytogenes from Raw Milk
Listeria monocytogenes has been reported as causative agent of foodborne disease which had gain public health concern as it can cause listeriosis in human especially in pregnant women, immunocompromised adults and infants. Various studies on molecular characterization of L. monocytogenes in foods...
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| Format: | Final Year Project Report / IMRAD |
| Language: | English English |
| Published: |
Universiti Malaysia Sarawak, UNIMAS
2013
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| Online Access: | http://ir.unimas.my/id/eprint/4451/ http://ir.unimas.my/id/eprint/4451/8/Gwo%20Rong%20%2824pgs%29.pdf http://ir.unimas.my/id/eprint/4451/10/Wong%20Gwo%20Rong.pdf |
| Summary: | Listeria monocytogenes has been reported as causative agent of foodborne disease which had gain public
health concern as it can cause listeriosis in human especially in pregnant women, immunocompromised
adults and infants. Various studies on molecular characterization of L. monocytogenes in foods had been
done in Malaysia such as chicken, beef and ready-to-eat foods. However, the study of L. monocytogenes in
raw milk is still not well established. The objectives of this study were to detect, isolate and characterize the
presence of L. monocytogenes from raw milk by using (GTG)5-PCR. Raw milk was collected from Kota
Samarahan and Kuching, Sarawak, and it was transported to laboratory for analysis. Samples were enriched
using Tryptone Soy Broth, then plated on PALCAM agar for isolation purposes. Next, the colonies formed
on PALCAM agar were confirmed by species-specific PCR using LM1 and LM2 specific primer for
detection of L. monocytogenes. The 234 bp hly gene was successfully amplified by PCR. Confirmed isolates
were characterized molecularly using (GTG)5-PCR method with 15-mer primers, GTG. (GTG)5-PCR results
were analysed using RAPDistance bioinformatics software. A dendrogram was successfully formed from
(GTG)5-PCR binding pattern and genetic distribution of L. monocytogenes was obtained. Based on the study,
14 positives L. monocytogenes were successfully isolated and characterized using (GTG)5-PCR from the total
of 120 isolates. Three clusters A, B and C showing the genetic distribution were formed from RAPDistance
based on (GTG)5-PCR binding patterns. In conclusion, 14 L. monocytogenes isolates were detected, isolated
and characterized into three clusters. |
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