Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types
The aim of this study was to develop a 16S-23S ribosomal deoxyribonucleic acid internal transcribed spacer (ITS) quantitative polymerase chain reaction (qPCR) assay for the early diagnosis and rapid screening of brucellosis. Blood, milk, and tissue samples were spiked with <i>B. abortus</i&...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-11-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/7/4/175 |
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| author | Rejoice Nyarku Ayesha Hassim Annelize Jonker Melvyn Quan |
| author_facet | Rejoice Nyarku Ayesha Hassim Annelize Jonker Melvyn Quan |
| author_sort | Rejoice Nyarku |
| collection | DOAJ |
| description | The aim of this study was to develop a 16S-23S ribosomal deoxyribonucleic acid internal transcribed spacer (ITS) quantitative polymerase chain reaction (qPCR) assay for the early diagnosis and rapid screening of brucellosis. Blood, milk, and tissue samples were spiked with <i>B. abortus</i> biovar 1 (B01988-18 strain) to determine the analytical sensitivity and specificity of the assay. The 95% limit of detection of the ITS qPCR assay was highest in tissue, followed by blood, then milk, i.e., 0.48, 4.43, and 15.18 bacteria/PCR reaction, respectively. The diagnostic performance of the assay was compared to the <i>Brucella</i> cell surface protein (BCSP) 31 qPCR assay and bacterial culture. Out of 56 aborted foetal tissue samples from bovine, ovine, and caprine, 33% (19/56) were positive for <i>Brucella</i> spp. The sensitivity and specificity of the ITS qPCR assay was 87% and 95% respectively, compared to 92% and 89% for the BCSP31 qPCR assay and 47% and 55% for bacterial culture, respectively. The assay was efficient, sensitive, and specific, making it a valuable tool in the early detection of the <i>Brucella</i> pathogen. |
| first_indexed | 2024-03-10T14:56:48Z |
| format | Article |
| id | doaj.art-4aa0a2ddbbaf4800b7dcd58947e34069 |
| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-03-10T14:56:48Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-4aa0a2ddbbaf4800b7dcd58947e340692023-11-20T20:30:26ZengMDPI AGVeterinary Sciences2306-73812020-11-017417510.3390/vetsci7040175Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen TypesRejoice Nyarku0Ayesha Hassim1Annelize Jonker2Melvyn Quan3Vectors and Vector-borne Diseases Research Programme, Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort 0110, South AfricaVectors and Vector-borne Diseases Research Programme, Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort 0110, South AfricaVectors and Vector-borne Diseases Research Programme, Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort 0110, South AfricaVectors and Vector-borne Diseases Research Programme, Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort 0110, South AfricaThe aim of this study was to develop a 16S-23S ribosomal deoxyribonucleic acid internal transcribed spacer (ITS) quantitative polymerase chain reaction (qPCR) assay for the early diagnosis and rapid screening of brucellosis. Blood, milk, and tissue samples were spiked with <i>B. abortus</i> biovar 1 (B01988-18 strain) to determine the analytical sensitivity and specificity of the assay. The 95% limit of detection of the ITS qPCR assay was highest in tissue, followed by blood, then milk, i.e., 0.48, 4.43, and 15.18 bacteria/PCR reaction, respectively. The diagnostic performance of the assay was compared to the <i>Brucella</i> cell surface protein (BCSP) 31 qPCR assay and bacterial culture. Out of 56 aborted foetal tissue samples from bovine, ovine, and caprine, 33% (19/56) were positive for <i>Brucella</i> spp. The sensitivity and specificity of the ITS qPCR assay was 87% and 95% respectively, compared to 92% and 89% for the BCSP31 qPCR assay and 47% and 55% for bacterial culture, respectively. The assay was efficient, sensitive, and specific, making it a valuable tool in the early detection of the <i>Brucella</i> pathogen.https://www.mdpi.com/2306-7381/7/4/175diagnosisqPCRbrucellosisbloodmilkabomasal fluid |
| spellingShingle | Rejoice Nyarku Ayesha Hassim Annelize Jonker Melvyn Quan Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types Veterinary Sciences diagnosis qPCR brucellosis blood milk abomasal fluid |
| title | Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types |
| title_full | Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types |
| title_fullStr | Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types |
| title_full_unstemmed | Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types |
| title_short | Development of a Genus-Specific <i>Brucella</i> Real-Time PCR Assay Targeting the 16S-23S rDNA Internal Transcribed Spacer from Different Specimen Types |
| title_sort | development of a genus specific i brucella i real time pcr assay targeting the 16s 23s rdna internal transcribed spacer from different specimen types |
| topic | diagnosis qPCR brucellosis blood milk abomasal fluid |
| url | https://www.mdpi.com/2306-7381/7/4/175 |
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