Sains Malaysiana 47(8)(2018): 1795–1800
http://dx.doi.org/10.17576/jsm-2018-4708-18
Detection of Leptospira Species in
Environmental Samples by Amplification of 16S rRNA and rpoβ Genes
(Pengesanan Spesies Leptospira dalam
Sampel Alam Sekitar melalui 16S rRNA dan Gen rpoβ Diperkuat)
NURUL FARHANA ZULKIFLI1, STEFFI JULAN WAN1, VASANTHA KUMARI NEELA2,
HUI
YEE
CHEE2, SITI
NORBAYA
MASRI2,
MAZEN
M.
JAMIL
AL-OBAIDI1
& MOHD NASIR MOHD DESA1,3*
1Department of Biomedical Science, Faculty
of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang,
Selangor Darul Ehsan, Malaysia
2Department of Medical Microbiology and
Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra
Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
3Laboratory of Halal Science Research, Halal Products
Research Institute, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor
Darul Ehsan, Malaysia
Diserahkan: 8 Febuari 2018/Diterima: 23
April 2018
ABSTRACT
This study attempted to identify and
determine distribution of Leptospira spp. in environmental samples
using 16S rRNA and rpoβ genes amplification. The samples
were collected from high risk areas in Selangor, Malaysia. A total of 105
environmental samples consisting of soil and water were subjected to direct DNA extraction
and PCR reaction. PCR products were analysed
using gel electrophoresis and subjected to sequence analysis. Thirteen out of
105 (12.38%) samples were amplified for 16S rRNA with an expected
amplicon size of 330 bp, while 50 out of 105 (47.62%) samples showed
amplification using rpoβ primers, but were not of expected size. Of
the 13 16S rRNA amplified samples, only 5 were identified as Leptospira in the gene sequence analysis and clustered under uncultured group via
phylogenetic tree. This study showed the DNA-based approach using PCR and
sequence analysis is able to detect the presence of Leptospira, although
environmental samples may contain diverse microbial populations that may
complicate the detection. Overall, the study suggested the importance of
surveillance for Leptospira from environmental samples.
Keywords: Environment; Leptospira;
rpoβ; Selangor; 16S
rRNA
ABSTRAK
Kajian ini dijalankan untuk mengenal
pasti dan menentukan penyebaran Leptospira spp. dalam sampel alam
sekitar menggunakan 16S rNA dan gen rpoβ diperkuat. Sampel
diperoleh daripada kawasan yang mempunyai risiko tinggi di Selangor, Malaysia.
Sampel keseluruhan adalah 105 yang terdiri daripada tanah dan air yang
tertakluk secara langsung kepada pengekstrakan DNA dan
reaksi PCR. Produk PCR dianalisis menggunakan
elektroforesis gel dan tertakluk kepada analisis urutan. Tiga belas sampel
daripada 105 (12.38%) menunjukkan amplifikasi untuk 16S rRNA dengan saiz
amplikon terjangka 330 bp, manakala 50 sampel daripada 105 (47.62%) menunjukkan
sampel amplikasi menggunakan pencetus rpoβ, tetapi tidak mengikut
saiz yang dijangka. Daripada 13 16S rRNA sampel diperkuat, hanya 5
dikenal pasti sebagai Leptospira dalam urutan gen analisis dan
berkelompok di bawah kumpulan tidak dibiak melalui pohon filogenetik. Kajian
ini menunjukkan pendekatan berasaskan DNA menggunakan PCR dan
analisis urutan dapat mengesan kehadiran Leptospira, walaupun sampel
alam sekitar berkemungkinan mengandungi pelbagai populasi mikrob yang boleh
merumitkan pengesanan. Secara keseluruhan, kajian ini mencadangkan kepentingan
pengawasan Leptospira daripada sampel alam sekitar.
Kata
kunci: Alam sekitar; Leptospira; rpoβ; Selangor;
16S rRNA
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*Pengarang untuk surat
menyurat; email: mnasir@upm.edu.my
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