Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei
Burkholderia mallei (B. mallei) is the etiological agent of glanders, a highly infectious and zoonotic disease of solipeds. Prompt and accurate detection of B. mallei and diagnosis of glanders is important for both, humans and animals. The present study was designed to evaluate the analytical sensit...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Kafkas University, Faculty of Veterinary Medicine
2017-03-01
|
| Series: | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| Subjects: | |
| Online Access: | https://vetdergikafkas.org/pdf.php?id=2045 |
| _version_ | 1797800552494530560 |
|---|---|
| author | KHAN I ALI S ELSCHNER M MERTENS K MELZER F AHMED H SAQIB M IJAZ M GWIDA M NEUBAUER H |
| author_facet | KHAN I ALI S ELSCHNER M MERTENS K MELZER F AHMED H SAQIB M IJAZ M GWIDA M NEUBAUER H |
| author_sort | KHAN I |
| collection | DOAJ |
| description | Burkholderia mallei (B. mallei) is the etiological agent of glanders, a highly infectious and zoonotic disease of solipeds. Prompt and accurate detection
of B. mallei and diagnosis of glanders is important for both, humans and animals. The present study was designed to evaluate the analytical sensitivity
and specificity of a commercially available one-step real-time (RT) PCR assay kit (Genekam Biotechnology AG, Germany) for the detection of B. mallei.
Furthermore, the comparative evaluation of the analytical sensitivity of an already published fliP RT-PCR protocol and this kit was made using a real-time
PCR (RT-PCR) platform: the Mx3000P TM (Stratagene®, Canada). Diagnostic parameters were assessed with a panel of 20 B. mallei, 20 B. pseudomallei, 15 B.
mallei related and 10 clinically relevant non-Burkholderia species. Using the one-step RT-PCR on the Mx3000PTM platform, the limit of detection (LOD) was
determined as 10 fg. Applying a modified fliP RT-PCR using the Mx3000P platform resulted in a LOD of 100 fg. Authors conclude that the one-step RT-PCR
kit is specific for DNA of B. mallei strains. The one-step RT-PCR assay kit is a robust, rapid, reliable, specific and sensitive tool with a fast turnaround time for
the specific identification and detection of B. mallei from culture material. |
| first_indexed | 2024-03-13T04:37:02Z |
| format | Article |
| id | doaj.art-23294569f5cc41b19c3ce5a9aba08a6c |
| institution | Directory Open Access Journal |
| issn | 1309-2251 |
| language | English |
| last_indexed | 2024-03-13T04:37:02Z |
| publishDate | 2017-03-01 |
| publisher | Kafkas University, Faculty of Veterinary Medicine |
| record_format | Article |
| series | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| spelling | doaj.art-23294569f5cc41b19c3ce5a9aba08a6c2023-06-19T06:46:03ZengKafkas University, Faculty of Veterinary MedicineKafkas Universitesi Veteriner Fakültesi Dergisi1309-22512017-03-0123231131710.9775/kvfd.2016.165182045Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia malleiKHAN I0ALI S1ELSCHNER M2MERTENS K3MELZER F4AHMED H5SAQIB M6IJAZ M7GWIDA M8NEUBAUER H9Section of Epidemiology and Public Health, University of Veterinary and Animal Sciences, Lahore, Sub-campus Jhang, PAKISTANDepartment of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore, PAKISTANFriedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit Naumburger Straße 96a, 07743, Jena, GERMANYFriedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit Naumburger Straße 96a, 07743, Jena, GERMANYFriedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit Naumburger Straße 96a, 07743, Jena, GERMANYDepartment of Biosciences, COMSATS Institute of İnformation Technology (CIIT), Park Road, Chakh Shahzad, Islamabad, PAKISTANDepartment of Clinical Medicine and Surgery, University of Agriculture, Faislabad, PAKISTANDepartment of Clinical Medicine and Surgery, University of Veterinary and Animal Sciences, Lahore, PAKISTANDepartment of Hygiene and Zoonoses, Faculty of Veterinary Medicine, Mansoura University, Mansoura 35516, EGYPTFriedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit Naumburger Straße 96a, 07743, Jena, GERMANYBurkholderia mallei (B. mallei) is the etiological agent of glanders, a highly infectious and zoonotic disease of solipeds. Prompt and accurate detection of B. mallei and diagnosis of glanders is important for both, humans and animals. The present study was designed to evaluate the analytical sensitivity and specificity of a commercially available one-step real-time (RT) PCR assay kit (Genekam Biotechnology AG, Germany) for the detection of B. mallei. Furthermore, the comparative evaluation of the analytical sensitivity of an already published fliP RT-PCR protocol and this kit was made using a real-time PCR (RT-PCR) platform: the Mx3000P TM (Stratagene®, Canada). Diagnostic parameters were assessed with a panel of 20 B. mallei, 20 B. pseudomallei, 15 B. mallei related and 10 clinically relevant non-Burkholderia species. Using the one-step RT-PCR on the Mx3000PTM platform, the limit of detection (LOD) was determined as 10 fg. Applying a modified fliP RT-PCR using the Mx3000P platform resulted in a LOD of 100 fg. Authors conclude that the one-step RT-PCR kit is specific for DNA of B. mallei strains. The one-step RT-PCR assay kit is a robust, rapid, reliable, specific and sensitive tool with a fast turnaround time for the specific identification and detection of B. mallei from culture material.https://vetdergikafkas.org/pdf.php?id=2045burkholderia malleianalytical specificityanalytical sensitivityone-step rt-pcr kit |
| spellingShingle | KHAN I ALI S ELSCHNER M MERTENS K MELZER F AHMED H SAQIB M IJAZ M GWIDA M NEUBAUER H Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei Kafkas Universitesi Veteriner Fakültesi Dergisi burkholderia mallei analytical specificity analytical sensitivity one-step rt-pcr kit |
| title | Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| title_full | Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| title_fullStr | Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| title_full_unstemmed | Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| title_short | Evaluation of analytical-sensitivity and -specificity of a commercially available one-step real-time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| title_sort | evaluation of analytical sensitivity and specificity of a commercially available one step real time polymerase chain reaction assay kit for the detection of burkholderia mallei |
| topic | burkholderia mallei analytical specificity analytical sensitivity one-step rt-pcr kit |
| url | https://vetdergikafkas.org/pdf.php?id=2045 |
| work_keys_str_mv | AT khani evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT alis evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT elschnerm evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT mertensk evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT melzerf evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT ahmedh evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT saqibm evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT ijazm evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT gwidam evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei AT neubauerh evaluationofanalyticalsensitivityandspecificityofacommerciallyavailableonesteprealtimepolymerasechainreactionassaykitforthedetectionofburkholderiamallei |