Detection of Brucella abortus by alkB and IS711 based primers
<font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>BACKGROUND<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wolters Kluwer Medknow Publications
2007-06-01
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| Series: | Journal of Research in Medical Sciences |
| Subjects: | |
| Online Access: | http://journals.mui.ac.ir/jrms/article/view/190 |
| _version_ | 1811281404096413696 |
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| author | Seyed Reza Hosseini Doust Zeinab Ahamdi Ali Ahamdi Massoud Hajia Morteza Izadi Ashraf Mohabati Mobarez |
| author_facet | Seyed Reza Hosseini Doust Zeinab Ahamdi Ali Ahamdi Massoud Hajia Morteza Izadi Ashraf Mohabati Mobarez |
| author_sort | Seyed Reza Hosseini Doust |
| collection | DOAJ |
| description | <font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>BACKGROUND<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">Brucellosis is a zoonotic disease, which involves both animals and human. Although the conventional methods have been widely used for its laboratory diagnosis, the PCR techniques have proved to be useful due to specificity, sensitivity and the rapidness. Various target sequences of brucella bacterium such as OMP2, 16s RNA and IS711 have been used for the primer designing.. All primer sets have shown different sensitivities and specificities. In present investigation, PCR protocol and primer designated based on IS711 and a fragment of chromosomal DNA all were optimized with standard genome and clinical samples.</font></p></font><p align="left"><strong><font face="TimesNewRoman,Bold" size="1">METHODS</font></strong><font face="TimesNewRoman" size="1">: </font><font face="TimesNewRoman" size="2">Numerous tissue samples (liver, kidney, lymph node, and uterus) were prepared and were cultured by the bacteriological standard methods along with the serology positive human samples. PCR protocol was optimized and the primer&#39;s sensitivity and the specificity were checked using pure genome of </font><em><font face="TimesNewRoman,Italic" size="2">B. abortus</font></em><font face="TimesNewRoman" size="2">. All samples were tested by the standard bacteriological methods. The samples were then subject to PCR amplification and the PCR product was confirmed using the RFLP technique.</font></p><font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>RESULTS<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">The culture results indicated a poor sensitivity as it was previously reported. The PCR product 157 bp was observed on the agarose gel indicating that significant number of clinical samples (human brucellosis cases) were positive by PCR but not by the culture method. Although </font><em><font face="TimesNewRoman,Italic" size="2">B. abortus </font></em><font face="TimesNewRoman" size="2">DNA was detected in all the culture positive veterinary specimens, some cross-reactions with close related bacteria were observed that might influence the interpretation of the results.</font></p></font><font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>CONCLUSIONS<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">The sensitivity of the present PCR protocol was significantly higher when alk B and IS711 based primers were used in compare to each of the alkB and IS711 based primers alone. More research will be needed to improve the specificity and sensitivity of the PCR protocol before recommending for routine laboratory works.</font></p></font><p align="left"><strong><font face="TimesNewRoman,Bold" size="1">KEY WORDS: </font></strong><font face="TimesNewRoman" size="2">Brucella, PCR, RFLP, brucellosis.</font></p> |
| first_indexed | 2024-04-13T01:33:14Z |
| format | Article |
| id | doaj.art-64e9146994d746709e5071a6f0a1f4e9 |
| institution | Directory Open Access Journal |
| issn | 1735-1995 1735-7136 |
| language | English |
| last_indexed | 2024-04-13T01:33:14Z |
| publishDate | 2007-06-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Journal of Research in Medical Sciences |
| spelling | doaj.art-64e9146994d746709e5071a6f0a1f4e92022-12-22T03:08:28ZengWolters Kluwer Medknow PublicationsJournal of Research in Medical Sciences1735-19951735-71362007-06-011226267Detection of Brucella abortus by alkB and IS711 based primersSeyed Reza Hosseini DoustZeinab AhamdiAli AhamdiMassoud HajiaMorteza IzadiAshraf Mohabati Mobarez<font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>BACKGROUND<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">Brucellosis is a zoonotic disease, which involves both animals and human. Although the conventional methods have been widely used for its laboratory diagnosis, the PCR techniques have proved to be useful due to specificity, sensitivity and the rapidness. Various target sequences of brucella bacterium such as OMP2, 16s RNA and IS711 have been used for the primer designing.. All primer sets have shown different sensitivities and specificities. In present investigation, PCR protocol and primer designated based on IS711 and a fragment of chromosomal DNA all were optimized with standard genome and clinical samples.</font></p></font><p align="left"><strong><font face="TimesNewRoman,Bold" size="1">METHODS</font></strong><font face="TimesNewRoman" size="1">: </font><font face="TimesNewRoman" size="2">Numerous tissue samples (liver, kidney, lymph node, and uterus) were prepared and were cultured by the bacteriological standard methods along with the serology positive human samples. PCR protocol was optimized and the primer&#39;s sensitivity and the specificity were checked using pure genome of </font><em><font face="TimesNewRoman,Italic" size="2">B. abortus</font></em><font face="TimesNewRoman" size="2">. All samples were tested by the standard bacteriological methods. The samples were then subject to PCR amplification and the PCR product was confirmed using the RFLP technique.</font></p><font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>RESULTS<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">The culture results indicated a poor sensitivity as it was previously reported. The PCR product 157 bp was observed on the agarose gel indicating that significant number of clinical samples (human brucellosis cases) were positive by PCR but not by the culture method. Although </font><em><font face="TimesNewRoman,Italic" size="2">B. abortus </font></em><font face="TimesNewRoman" size="2">DNA was detected in all the culture positive veterinary specimens, some cross-reactions with close related bacteria were observed that might influence the interpretation of the results.</font></p></font><font face="TimesNewRoman,Bold" size="1"><p align="left"><strong>CONCLUSIONS<font face="TimesNewRoman" size="1">: </font></strong><font face="TimesNewRoman" size="2">The sensitivity of the present PCR protocol was significantly higher when alk B and IS711 based primers were used in compare to each of the alkB and IS711 based primers alone. More research will be needed to improve the specificity and sensitivity of the PCR protocol before recommending for routine laboratory works.</font></p></font><p align="left"><strong><font face="TimesNewRoman,Bold" size="1">KEY WORDS: </font></strong><font face="TimesNewRoman" size="2">Brucella, PCR, RFLP, brucellosis.</font></p>http://journals.mui.ac.ir/jrms/article/view/190Brucella, PCR, RFLP, brucellosis. |
| spellingShingle | Seyed Reza Hosseini Doust Zeinab Ahamdi Ali Ahamdi Massoud Hajia Morteza Izadi Ashraf Mohabati Mobarez Detection of Brucella abortus by alkB and IS711 based primers Journal of Research in Medical Sciences Brucella, PCR, RFLP, brucellosis. |
| title | Detection of Brucella abortus by alkB and IS711 based primers |
| title_full | Detection of Brucella abortus by alkB and IS711 based primers |
| title_fullStr | Detection of Brucella abortus by alkB and IS711 based primers |
| title_full_unstemmed | Detection of Brucella abortus by alkB and IS711 based primers |
| title_short | Detection of Brucella abortus by alkB and IS711 based primers |
| title_sort | detection of brucella abortus by alkb and is711 based primers |
| topic | Brucella, PCR, RFLP, brucellosis. |
| url | http://journals.mui.ac.ir/jrms/article/view/190 |
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