Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates
Introduction: Streptococcus suis is a zoonotic gram-positive cocci causing systemic infection and severe acute meningitis in humans. Infection is involved in the exposure of contaminated pigs and raw pork. Endemic usually occurs in northern area of Thailand where people eat raw pork-derived products...
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| Format: | Article |
| Language: | English |
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Chaing Mai University
2016-05-01
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| Series: | Journal of Associated Medical Sciences |
| Subjects: | |
| Online Access: | https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/59892 |
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| author | Sorasak Intorasoot Amornrat Intorasoot Supaporn Limpaisarn Nutsuda Kojorla Santhasiri Orrapin Manlika Wanmakok Banyong Khantawa |
| author_facet | Sorasak Intorasoot Amornrat Intorasoot Supaporn Limpaisarn Nutsuda Kojorla Santhasiri Orrapin Manlika Wanmakok Banyong Khantawa |
| author_sort | Sorasak Intorasoot |
| collection | DOAJ |
| description | Introduction: Streptococcus suis is a zoonotic gram-positive cocci causing systemic infection and severe acute meningitis in humans. Infection is involved in the exposure of contaminated pigs and raw pork. Endemic usually occurs in northern area of Thailand where people eat raw pork-derived products. Due to lacking of effective vaccines, an early diagnosis of S. suis infection is extremely important for disease control and prevention.
Objective: To develop a rapid and high sensitive loop-mediated isothermal amplification (LAMP) technique for investigation of S. suis.
Materials and methods: Based on conserved sequences, glutamate dehydrogenase gene of S. suis was used as a target for LAMP primers design. The detection limit was compared with a conventional PCR. Thirty-four serotypes consisting of serotype 1-31, 32, 34 and 1/2 of S. suis were examined with the developed method. Specificity determination was tested with several blood-borne bacteria including other alpha and beta hemolytic Streptococcus spp. In addition, 25 clinical isolates and 30 positive hemocultures were resolved by established LAMP assay.
Results: LAMP assay exhibited approximately 1,000 times more sensitive than the conventional PCR with a final bacterial load of approximately 12 colony forming unit (CFU). Thirty-four serotypes of S. suis were tested and 24 of those – serotype 2-4, 6-12, 14, 15, 17-19, 21, 24-26, 28-31 and 1/2 could be detected by LAMP technique. The assay demonstrated a high specificity without cross-reactivity to other bacteria. In addition, 25 clinical isolates and 2 out of 30 cases from positive hemocultures were successfully amplified by this method.
Conclusion: LAMP assay developed is rapid and sensitive, and can be used for routine diagnosis of various serotypes of S. suis infection in clinical specimens.
Bull Chiang Mai Assoc Med Sci 2016; 49(2): 207-217. Doi: 10.14456/jams.2016.23 |
| first_indexed | 2024-04-13T17:14:37Z |
| format | Article |
| id | doaj.art-688623290a3c4b099ddf46a1f138c12f |
| institution | Directory Open Access Journal |
| issn | 2539-6056 2539-6056 |
| language | English |
| last_indexed | 2024-04-13T17:14:37Z |
| publishDate | 2016-05-01 |
| publisher | Chaing Mai University |
| record_format | Article |
| series | Journal of Associated Medical Sciences |
| spelling | doaj.art-688623290a3c4b099ddf46a1f138c12f2022-12-22T02:38:09ZengChaing Mai UniversityJournal of Associated Medical Sciences2539-60562539-60562016-05-0149220720759892Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolatesSorasak Intorasoot0Amornrat Intorasoot1Supaporn Limpaisarn2Nutsuda Kojorla3Santhasiri Orrapin4Manlika Wanmakok5Banyong Khantawa61Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai University1Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University 2Department of Microbiology, Faculty of Medicine, Chiang Mai University 3Clinical Microbiology Section, Central Diagnostic Laboratory, Maharaj Nakorn Chiang Mai Hospital, Faculty of Medicine, Chiang Mai UniversityIntroduction: Streptococcus suis is a zoonotic gram-positive cocci causing systemic infection and severe acute meningitis in humans. Infection is involved in the exposure of contaminated pigs and raw pork. Endemic usually occurs in northern area of Thailand where people eat raw pork-derived products. Due to lacking of effective vaccines, an early diagnosis of S. suis infection is extremely important for disease control and prevention. Objective: To develop a rapid and high sensitive loop-mediated isothermal amplification (LAMP) technique for investigation of S. suis. Materials and methods: Based on conserved sequences, glutamate dehydrogenase gene of S. suis was used as a target for LAMP primers design. The detection limit was compared with a conventional PCR. Thirty-four serotypes consisting of serotype 1-31, 32, 34 and 1/2 of S. suis were examined with the developed method. Specificity determination was tested with several blood-borne bacteria including other alpha and beta hemolytic Streptococcus spp. In addition, 25 clinical isolates and 30 positive hemocultures were resolved by established LAMP assay. Results: LAMP assay exhibited approximately 1,000 times more sensitive than the conventional PCR with a final bacterial load of approximately 12 colony forming unit (CFU). Thirty-four serotypes of S. suis were tested and 24 of those – serotype 2-4, 6-12, 14, 15, 17-19, 21, 24-26, 28-31 and 1/2 could be detected by LAMP technique. The assay demonstrated a high specificity without cross-reactivity to other bacteria. In addition, 25 clinical isolates and 2 out of 30 cases from positive hemocultures were successfully amplified by this method. Conclusion: LAMP assay developed is rapid and sensitive, and can be used for routine diagnosis of various serotypes of S. suis infection in clinical specimens. Bull Chiang Mai Assoc Med Sci 2016; 49(2): 207-217. Doi: 10.14456/jams.2016.23https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/59892Streptococcus suisglutamate dehydrogenaseLAMPhemoculturePCR |
| spellingShingle | Sorasak Intorasoot Amornrat Intorasoot Supaporn Limpaisarn Nutsuda Kojorla Santhasiri Orrapin Manlika Wanmakok Banyong Khantawa Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates Journal of Associated Medical Sciences Streptococcus suis glutamate dehydrogenase LAMP hemoculture PCR |
| title | Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates |
| title_full | Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates |
| title_fullStr | Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates |
| title_full_unstemmed | Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates |
| title_short | Loop-mediated isothermal amplification for rapid detection of Streptococcus suis in hemoculture and clinical isolates |
| title_sort | loop mediated isothermal amplification for rapid detection of streptococcus suis in hemoculture and clinical isolates |
| topic | Streptococcus suis glutamate dehydrogenase LAMP hemoculture PCR |
| url | https://www.tci-thaijo.org/index.php/bulletinAMS/article/view/59892 |
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