Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent.
Glanders is a highly contagious and potentially serious disease caused by Burkholderia mallei, a Tier 1 select agent. In this study, we raised a monoclonal antibody (mAb) against the lipopolysaccharide (LPS) of B. mallei and developed a competitive enzyme-linked immunosorbent assay (cELISA) for B. m...
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ftdoajarticles:oai:doaj.org/article:f5363875e0f8494c88ffdba6ccb584e7 2023-05-15T15:11:10+02:00 Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. Ulrich Wernery Elaine Chan Rekha Raghavan Jade L L Teng Ginu Syriac Sing-Yung Siu Marina Joseph Man-Lung Yeung Lilong Jia Jian-Piao Cai Tsz-Ho Chiu Susanna K P Lau Patrick C Y Woo 2021-12-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0010007 https://doaj.org/article/f5363875e0f8494c88ffdba6ccb584e7 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0010007 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0010007 https://doaj.org/article/f5363875e0f8494c88ffdba6ccb584e7 PLoS Neglected Tropical Diseases, Vol 15, Iss 12, p e0010007 (2021) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2021 ftdoajarticles https://doi.org/10.1371/journal.pntd.0010007 2022-12-31T15:08:13Z Glanders is a highly contagious and potentially serious disease caused by Burkholderia mallei, a Tier 1 select agent. In this study, we raised a monoclonal antibody (mAb) against the lipopolysaccharide (LPS) of B. mallei and developed a competitive enzyme-linked immunosorbent assay (cELISA) for B. mallei infection. Using the titrated optimal conditions of B. mallei-LPS (2 ng) for microtiter plate coating, sample serum dilution at 1:20 and 3.5 ng/μL anti-LPS mAb B5, the cutoff value of the cELISA was determined using serum samples from 136 glanders-free seronegative horses in Hong Kong. All calculated percentage inhibition (PI) values from these seronegative samples were below 39.6% inhibition (1.5 standard deviations above mean PI) and was used as the cutoff value. The diagnostic sensitivity of the developed LPS-based cELISA was first evaluated using sera from donkeys and mice inoculated with B. mallei. An increasing trend of PI values above the defined cELISA cutoff observed in the donkey and mouse sera suggested positive detection of anti-LPS antibodies. The sensitivity and specificity of the LPS-based cELISA was further evaluated using 31 serologically positive horse sera from glanders outbreaks in Bahrain and Kuwait, of which 30 were tested positive by the cELISA; and 21 seronegative horse sera and 20 seronegative donkey sera from Dubai, of which all were tested negative by the cELISA. A cELISA with high sensitivity (97.2%) and specificity (100%) for the detection of B. mallei antibodies in different animals was developed. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 15 12 e0010007 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Ulrich Wernery Elaine Chan Rekha Raghavan Jade L L Teng Ginu Syriac Sing-Yung Siu Marina Joseph Man-Lung Yeung Lilong Jia Jian-Piao Cai Tsz-Ho Chiu Susanna K P Lau Patrick C Y Woo Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Glanders is a highly contagious and potentially serious disease caused by Burkholderia mallei, a Tier 1 select agent. In this study, we raised a monoclonal antibody (mAb) against the lipopolysaccharide (LPS) of B. mallei and developed a competitive enzyme-linked immunosorbent assay (cELISA) for B. mallei infection. Using the titrated optimal conditions of B. mallei-LPS (2 ng) for microtiter plate coating, sample serum dilution at 1:20 and 3.5 ng/μL anti-LPS mAb B5, the cutoff value of the cELISA was determined using serum samples from 136 glanders-free seronegative horses in Hong Kong. All calculated percentage inhibition (PI) values from these seronegative samples were below 39.6% inhibition (1.5 standard deviations above mean PI) and was used as the cutoff value. The diagnostic sensitivity of the developed LPS-based cELISA was first evaluated using sera from donkeys and mice inoculated with B. mallei. An increasing trend of PI values above the defined cELISA cutoff observed in the donkey and mouse sera suggested positive detection of anti-LPS antibodies. The sensitivity and specificity of the LPS-based cELISA was further evaluated using 31 serologically positive horse sera from glanders outbreaks in Bahrain and Kuwait, of which 30 were tested positive by the cELISA; and 21 seronegative horse sera and 20 seronegative donkey sera from Dubai, of which all were tested negative by the cELISA. A cELISA with high sensitivity (97.2%) and specificity (100%) for the detection of B. mallei antibodies in different animals was developed. |
format |
Article in Journal/Newspaper |
author |
Ulrich Wernery Elaine Chan Rekha Raghavan Jade L L Teng Ginu Syriac Sing-Yung Siu Marina Joseph Man-Lung Yeung Lilong Jia Jian-Piao Cai Tsz-Ho Chiu Susanna K P Lau Patrick C Y Woo |
author_facet |
Ulrich Wernery Elaine Chan Rekha Raghavan Jade L L Teng Ginu Syriac Sing-Yung Siu Marina Joseph Man-Lung Yeung Lilong Jia Jian-Piao Cai Tsz-Ho Chiu Susanna K P Lau Patrick C Y Woo |
author_sort |
Ulrich Wernery |
title |
Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
title_short |
Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
title_full |
Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
title_fullStr |
Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
title_full_unstemmed |
Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent. |
title_sort |
development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of burkholderia mallei, a tier 1 select agent. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2021 |
url |
https://doi.org/10.1371/journal.pntd.0010007 https://doaj.org/article/f5363875e0f8494c88ffdba6ccb584e7 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 15, Iss 12, p e0010007 (2021) |
op_relation |
https://doi.org/10.1371/journal.pntd.0010007 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0010007 https://doaj.org/article/f5363875e0f8494c88ffdba6ccb584e7 |
op_doi |
https://doi.org/10.1371/journal.pntd.0010007 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
15 |
container_issue |
12 |
container_start_page |
e0010007 |
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1766342074228015104 |