APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION (m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN Mycobacterium bovis PADA DOMBA
Tuberculosis (TB) is an important zoonotic disease which until now is a major problem in both humans and animals. Early diagnosis of TB is a very important issue considering TB disease can be transmitted through the air, so that rapid and accurate diagnosis is a key in preventing the spread of this...
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2013
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id-ugm-repo.1231662016-03-04T08:44:48Z https://repository.ugm.ac.id/123166/ APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION (m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN Mycobacterium bovis PADA DOMBA , Yohanes Timbun Raja Mangihut Ronael Simarmata , Prof. Dr. drh. Ida Tjahajati, MP. ETD Tuberculosis (TB) is an important zoonotic disease which until now is a major problem in both humans and animals. Early diagnosis of TB is a very important issue considering TB disease can be transmitted through the air, so that rapid and accurate diagnosis is a key in preventing the spread of this disease. The fact of TB in cattle in the field as a potential source of transmission to humans, encouraging the development of methods for rapid and accurate diagnosis. The m-PCR method which can detect more than one type of bacteria at a time, enabling the construction of TB diagnosis kit which can simultaneously detect the presence of M. tuberculosis and M.bovis which is the cause of tuberculosis that is harmful to animals and humans. The purpose of this research is the application of m-PCR method for detection of M. tuberculosis and M.bovis in sheep with the specific primers that can differentiate the DNA of M. tuberculosis and M. bovis. The samples used were obtained from pulmonary and limfoglandula organs of sheep (ovis aries) with suspect TB from Yogyakarta. Isolation of DNA were conducted using PureLink TM The m-PCR results from samples of lung and limfoglandula of the suspect sheep showed 4 sheep infected by M tuberculosis and M.bovis. Two isolated DNA showed at position of 168 bp (positive M.bovis), and 2 isolates were at 297 bp (positive M. tuberculosis). Sequences analysis results showed a highly similarity (90% -100%) with others nucleotide sequences of Genomic DNA kit Invitrogen production by following the manufacturer's procedure. Isolated DNA then were used as template in the m-PCR amplification process using forward primer 5'-TTCCGAATCCCTTGTGA-3', coded CSB1, reverse primer 5'- GGAGAGCGCCGTTGTA-3', coded CSB2 (M.bovis) and reverse primer 5'- AGTCGCGTGGCTTCTCTTTTA-3', coded CSB3 (M.tuberculosis). The PCR amplification product then were reconfirmed by agarose gel electrophoresis as positive result showed at the band of 297 bp (M.tuberculosis) and 168 bp (M.bovis). M.bovis BCG str. Korea and M. tuberculosis H37Rv complete genomes in GenBank. The result were reinforced by the growth of bacterial colonies on ogawa media. From the whole analysis, this study conclude that the m-PCR method using specific primers (CSB1, CSB2, CSB3) can be applied for detection of M. tuberculosis and M. bovis on sheep. The TB disease in sheep can be caused by M.tuberculosis, besides M.bovis and M.caprae. [Yogyakarta] : Universitas Gadjah Mada 2013 Thesis NonPeerReviewed , Yohanes Timbun Raja Mangihut Ronael Simarmata and , Prof. Dr. drh. Ida Tjahajati, MP. (2013) APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION (m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN Mycobacterium bovis PADA DOMBA. UNSPECIFIED thesis, UNSPECIFIED. http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=63277 |
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ETD , Yohanes Timbun Raja Mangihut Ronael Simarmata , Prof. Dr. drh. Ida Tjahajati, MP. APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION (m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN Mycobacterium bovis PADA DOMBA |
description |
Tuberculosis (TB) is an important zoonotic disease which until now is
a major problem in both humans and animals. Early diagnosis of TB is a very
important issue considering TB disease can be transmitted through the air, so
that rapid and accurate diagnosis is a key in preventing the spread of this
disease. The fact of TB in cattle in the field as a potential source of
transmission to humans, encouraging the development of methods for rapid
and accurate diagnosis. The m-PCR method which can detect more than one
type of bacteria at a time, enabling the construction of TB diagnosis kit which
can simultaneously detect the presence of M. tuberculosis and M.bovis which
is the cause of tuberculosis that is harmful to animals and humans. The
purpose of this research is the application of m-PCR method for detection of
M. tuberculosis and M.bovis in sheep with the specific primers that can
differentiate the DNA of M. tuberculosis and M. bovis.
The samples used were obtained from pulmonary and limfoglandula
organs of sheep (ovis aries) with suspect TB from Yogyakarta. Isolation of
DNA were conducted using PureLink TM
The m-PCR results from samples of lung and limfoglandula of the
suspect sheep showed 4 sheep infected by M tuberculosis and M.bovis. Two
isolated DNA showed at position of 168 bp (positive M.bovis), and 2 isolates
were at 297 bp (positive M. tuberculosis). Sequences analysis results showed
a highly similarity (90% -100%) with others nucleotide sequences of
Genomic DNA kit Invitrogen
production by following the manufacturer's procedure. Isolated DNA then
were used as template in the m-PCR amplification process using forward
primer 5'-TTCCGAATCCCTTGTGA-3', coded CSB1, reverse primer 5'-
GGAGAGCGCCGTTGTA-3', coded CSB2 (M.bovis) and reverse primer 5'-
AGTCGCGTGGCTTCTCTTTTA-3', coded CSB3 (M.tuberculosis). The
PCR amplification product then were reconfirmed by agarose gel
electrophoresis as positive result showed at the band of 297 bp
(M.tuberculosis) and 168 bp (M.bovis).
M.bovis
BCG str. Korea and M. tuberculosis H37Rv complete genomes in GenBank.
The result were reinforced by the growth of bacterial colonies on ogawa
media. From the whole analysis, this study conclude that the m-PCR method
using specific primers (CSB1, CSB2, CSB3) can be applied for detection of
M. tuberculosis and M. bovis on sheep. The TB disease in sheep can be
caused by M.tuberculosis, besides M.bovis and M.caprae. |
format |
Theses and Dissertations NonPeerReviewed |
author |
, Yohanes Timbun Raja Mangihut Ronael Simarmata , Prof. Dr. drh. Ida Tjahajati, MP. |
author_facet |
, Yohanes Timbun Raja Mangihut Ronael Simarmata , Prof. Dr. drh. Ida Tjahajati, MP. |
author_sort |
, Yohanes Timbun Raja Mangihut Ronael Simarmata |
title |
APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION
(m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN
Mycobacterium bovis PADA DOMBA |
title_short |
APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION
(m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN
Mycobacterium bovis PADA DOMBA |
title_full |
APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION
(m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN
Mycobacterium bovis PADA DOMBA |
title_fullStr |
APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION
(m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN
Mycobacterium bovis PADA DOMBA |
title_full_unstemmed |
APLIKASI MULTIPLEX POLYMERASE CHAIN REACTION
(m-PCR) UNTUK DETEKSI Mycobacterium tuberculosis DAN
Mycobacterium bovis PADA DOMBA |
title_sort |
aplikasi multiplex polymerase chain reaction
(m-pcr) untuk deteksi mycobacterium tuberculosis dan
mycobacterium bovis pada domba |
publisher |
[Yogyakarta] : Universitas Gadjah Mada |
publishDate |
2013 |
url |
https://repository.ugm.ac.id/123166/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=63277 |
_version_ |
1681231840089735168 |