Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide
Brucellosis is an important zoonotic disease with considerable impacts on human and animal health. Brucella abortus strain RB51 vaccine is used for prevention of bovine brucellosis in Iran. Due to strain roughness, available serological tests cannot detect vaccinated animals. Detection of serologica...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria
2019-06-01
|
| Series: | Bulgarian Journal of Veterinary Medicine |
| Subjects: | |
| Online Access: | http://tru.uni-sz.bg/bjvm/BJVM%20June%202019%20p.152-162.pdf |
| _version_ | 1818023642503577600 |
|---|---|
| author | A. Kalantari T. Naserpour Farivar A. Peymani M. Aslanimehr R. Bagheri Nejad |
| author_facet | A. Kalantari T. Naserpour Farivar A. Peymani M. Aslanimehr R. Bagheri Nejad |
| author_sort | A. Kalantari |
| collection | DOAJ |
| description | Brucellosis is an important zoonotic disease with considerable impacts on human and animal health. Brucella abortus strain RB51 vaccine is used for prevention of bovine brucellosis in Iran. Due to strain roughness, available serological tests cannot detect vaccinated animals. Detection of serological responses to the vaccine is important to monitor accurate vaccination implementation. Rough lipopolysaccharide (RLPS) of RB51 strain was extracted and characterised to develop serological tests for diagnosis of vaccinated animals. RLPS was extracted using phenol-chloroform-petroleum ether and evaluated by limulus amebocyte lysate (LAL) assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and agar gel immunodiffusion (AGID). According to our results, the extracted RLPS caused positive reaction in LAL assay. In SDS-PAGE, a band with a molecular weight around 14 kDa was identified after specific staining using silver nitrate. Double AGID of the RLPS with a hyperimmune serum resulted in a precipitation line formation. Our study showed that the method can be successfully used to extract RLPS from Brucella abortus strain RB51 as confirmed by LAL assay, PAGE and AGID. |
| first_indexed | 2024-12-10T03:47:34Z |
| format | Article |
| id | doaj.art-33d22ce3921048d29c9e8cede30742b9 |
| institution | Directory Open Access Journal |
| issn | 1311-1477 1313-3543 |
| language | English |
| last_indexed | 2024-12-10T03:47:34Z |
| publishDate | 2019-06-01 |
| publisher | Faculty of Veterinary Medicine, Trakia University, Stara Zagora, Bulgaria |
| record_format | Article |
| series | Bulgarian Journal of Veterinary Medicine |
| spelling | doaj.art-33d22ce3921048d29c9e8cede30742b92022-12-22T02:03:21ZengFaculty of Veterinary Medicine, Trakia University, Stara Zagora, BulgariaBulgarian Journal of Veterinary Medicine1311-14771313-35432019-06-0122215216210.15547/bjvm.2050Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharideA. Kalantari0T. Naserpour Farivar1A. Peymani2M. Aslanimehr 3R. Bagheri Nejad4Cellular & Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IranCellular & Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IranCellular & Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IranCellular & Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, IranBrucellosis Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, IranBrucellosis is an important zoonotic disease with considerable impacts on human and animal health. Brucella abortus strain RB51 vaccine is used for prevention of bovine brucellosis in Iran. Due to strain roughness, available serological tests cannot detect vaccinated animals. Detection of serological responses to the vaccine is important to monitor accurate vaccination implementation. Rough lipopolysaccharide (RLPS) of RB51 strain was extracted and characterised to develop serological tests for diagnosis of vaccinated animals. RLPS was extracted using phenol-chloroform-petroleum ether and evaluated by limulus amebocyte lysate (LAL) assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and agar gel immunodiffusion (AGID). According to our results, the extracted RLPS caused positive reaction in LAL assay. In SDS-PAGE, a band with a molecular weight around 14 kDa was identified after specific staining using silver nitrate. Double AGID of the RLPS with a hyperimmune serum resulted in a precipitation line formation. Our study showed that the method can be successfully used to extract RLPS from Brucella abortus strain RB51 as confirmed by LAL assay, PAGE and AGID.http://tru.uni-sz.bg/bjvm/BJVM%20June%202019%20p.152-162.pdf: brucellosisRB51 vaccinerough lipopolysaccharide |
| spellingShingle | A. Kalantari T. Naserpour Farivar A. Peymani M. Aslanimehr R. Bagheri Nejad Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide Bulgarian Journal of Veterinary Medicine : brucellosis RB51 vaccine rough lipopolysaccharide |
| title | Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide |
| title_full | Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide |
| title_fullStr | Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide |
| title_full_unstemmed | Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide |
| title_short | Extraction and characterisation of Brucella abortus strain RB51 rough lipopolysaccharide |
| title_sort | extraction and characterisation of brucella abortus strain rb51 rough lipopolysaccharide |
| topic | : brucellosis RB51 vaccine rough lipopolysaccharide |
| url | http://tru.uni-sz.bg/bjvm/BJVM%20June%202019%20p.152-162.pdf |
| work_keys_str_mv | AT akalantari extractionandcharacterisationofbrucellaabortusstrainrb51roughlipopolysaccharide AT tnaserpourfarivar extractionandcharacterisationofbrucellaabortusstrainrb51roughlipopolysaccharide AT apeymani extractionandcharacterisationofbrucellaabortusstrainrb51roughlipopolysaccharide AT maslanimehr extractionandcharacterisationofbrucellaabortusstrainrb51roughlipopolysaccharide AT rbagherinejad extractionandcharacterisationofbrucellaabortusstrainrb51roughlipopolysaccharide |