Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish
Background and Objective: Streptococcosis/lactococcosis is the cause of high morbidity and mortality in aquaculture sector and to date a number of species of Streptococcus and Lactococcus genera including S. iniae, S. agalactiae, S. dysagalactiae, S. parauberis, S. feacalis, L. garvieae and L. lacti...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2012-06-01
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| Series: | Iranian Journal of Microbiology |
| Subjects: | |
| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/709 |
| _version_ | 1818368350707777536 |
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| author | M Soltani E Pirali P Shayan B Eckert S Rouholahi N Sadr-Shirazi |
| author_facet | M Soltani E Pirali P Shayan B Eckert S Rouholahi N Sadr-Shirazi |
| author_sort | M Soltani |
| collection | DOAJ |
| description | Background and Objective: Streptococcosis/lactococcosis is the cause of high morbidity and mortality in aquaculture sector and to date a number of species of Streptococcus and Lactococcus genera including S. iniae, S. agalactiae, S. dysagalactiae, S. parauberis, S. feacalis, L. garvieae and L. lactis have been discriminated as the cause of disease in aquatic animals. Despite the use of diagnostic molecular methods for each of these bacterial species, no data is available on a suitable, rapid and simple simultaneous detection tool for these pathogens. This paper describes a simultaneous detection method which is PCR based on a reverse line blot (RLB) for rapid detection and differentiation of four species of genera of Streptococcus and Lactococcus genera consisting of S. iniae, S. agalactiae, S. parauberis and L. garvieae the most important agents of the disease in fish.
Materials and Methods: A reverse line blot (RLB) assay was developed for the simultaneously identification of four species of Streptococcus/lactococcusconsisting of S. iniae, S. parauberis, S. agalactiaeand Lactococcusgarvieae. The assay employs one set of primer pair for specific amplification of the 16S rRNA gene. These were designed based on the nucleotide sequences of 16S rRNA gene sharing a homology region with Streptococcus spp. and Lactococcus spp. DNA was extracted from the pure bacterial colonies and amplified. A membrane was prepared with specific oligonucleotide for each bacterial species. PCR products were then hybridized to a membrane.
Results and Conclusion: The amplification resulted in PCR product of 241 bp in length. No cross-reactions were observed between any of the tested bacterial species, and mixed DNAs from these four bacterial species were correctly identified. This RLB method is a suitable technique for a simultaneous detection of these species of bacterial fish pathogens that are some of the main causes of streptococcal/lactococcal infections in both freshwater and marine aquatic animals, and so we recommend its use for integrated epidemiological monitoring of streptococcosis/lactococcis in aquaculture industry. |
| first_indexed | 2024-12-13T23:06:33Z |
| format | Article |
| id | doaj.art-8be1991e87cf4cd1b5cd1732e97fbba4 |
| institution | Directory Open Access Journal |
| issn | 2008-3289 2008-4447 |
| language | English |
| last_indexed | 2024-12-13T23:06:33Z |
| publishDate | 2012-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Microbiology |
| spelling | doaj.art-8be1991e87cf4cd1b5cd1732e97fbba42022-12-21T23:28:16ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472012-06-0142Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fishM Soltani0E Pirali1P Shayan2B Eckert3S Rouholahi4N Sadr-Shirazi5Department of aquatic animal health, faculty of veterinary medicine, university of Tehran, Tehran, Iran.Department of aquatic animal health, faculty of veterinary medicine, university of Tehran, Tehran, Iran.Department of parasitology, faculty of veterinary medicine, university of Tehran, Tehran, Iran.Investigation institute of molecular biology and system transfer (MBST), Tehran, Iran.Department of aquatic animal health, faculty of veterinary medicine, university of Tehran, Tehran, Iran.Investigation institute of molecular biology and system transfer (MBST), Tehran, Iran.Background and Objective: Streptococcosis/lactococcosis is the cause of high morbidity and mortality in aquaculture sector and to date a number of species of Streptococcus and Lactococcus genera including S. iniae, S. agalactiae, S. dysagalactiae, S. parauberis, S. feacalis, L. garvieae and L. lactis have been discriminated as the cause of disease in aquatic animals. Despite the use of diagnostic molecular methods for each of these bacterial species, no data is available on a suitable, rapid and simple simultaneous detection tool for these pathogens. This paper describes a simultaneous detection method which is PCR based on a reverse line blot (RLB) for rapid detection and differentiation of four species of genera of Streptococcus and Lactococcus genera consisting of S. iniae, S. agalactiae, S. parauberis and L. garvieae the most important agents of the disease in fish. Materials and Methods: A reverse line blot (RLB) assay was developed for the simultaneously identification of four species of Streptococcus/lactococcusconsisting of S. iniae, S. parauberis, S. agalactiaeand Lactococcusgarvieae. The assay employs one set of primer pair for specific amplification of the 16S rRNA gene. These were designed based on the nucleotide sequences of 16S rRNA gene sharing a homology region with Streptococcus spp. and Lactococcus spp. DNA was extracted from the pure bacterial colonies and amplified. A membrane was prepared with specific oligonucleotide for each bacterial species. PCR products were then hybridized to a membrane. Results and Conclusion: The amplification resulted in PCR product of 241 bp in length. No cross-reactions were observed between any of the tested bacterial species, and mixed DNAs from these four bacterial species were correctly identified. This RLB method is a suitable technique for a simultaneous detection of these species of bacterial fish pathogens that are some of the main causes of streptococcal/lactococcal infections in both freshwater and marine aquatic animals, and so we recommend its use for integrated epidemiological monitoring of streptococcosis/lactococcis in aquaculture industry.https://ijm.tums.ac.ir/index.php/ijm/article/view/709L. garviaeReverse line blot (RLB)bS. agalactiaeS. iniaeS. parauberis |
| spellingShingle | M Soltani E Pirali P Shayan B Eckert S Rouholahi N Sadr-Shirazi Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish Iranian Journal of Microbiology L. garviae Reverse line blot (RLB) bS. agalactiae S. iniae S. parauberis |
| title | Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish |
| title_full | Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish |
| title_fullStr | Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish |
| title_full_unstemmed | Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish |
| title_short | Development of a Reverse Line Blot Hybridization method for Detection of some Streptococcal/Lactococcal Species, the causative agents of Zoonotic Streptococosis/Lactococosis in farmed fish |
| title_sort | development of a reverse line blot hybridization method for detection of some streptococcal lactococcal species the causative agents of zoonotic streptococosis lactococosis in farmed fish |
| topic | L. garviae Reverse line blot (RLB) bS. agalactiae S. iniae S. parauberis |
| url | https://ijm.tums.ac.ir/index.php/ijm/article/view/709 |
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