A genetic variant of
            <i>Burkholderia mallei</i>
            detected in Kuwait: Consequences for the PCR diagnosis of glanders

Laroucau, Karine; Aaziz, Rachid; Vorimore, Fabien; Varghese, Koshy; Deshayes, Thomas; Bertin, Claire; Delannoy, Sabine; Sami, A.; Batel, Maha Al; Shorbagy, Mamdouh El; Almutawaa, Khaled A W; Alanezi, Saad J; Alazemi, Yousef S N; Guernier, Vanina; Wernery, Ulrich

doi:10.1111/tbed.13777

Cited by 16 publications

(13 citation statements)

References 25 publications

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“…Thorough genotype characterization of B. mallei strains would not only help to elucidate the origin of Pakistani strains, but also the detection and surveillance of glanders worldwide. Laroucau et al [36] reported that in two horses that were serologically tested and found to be positive for glanders, the routine PCR with tissue targeting the fliP-IS407A gene did not give a positive result, probably due to genetic variation in the infecting strains. Solely by applying more extensive methods, like SNP and MLST typing, the serological results could be confirmed.…”

Section: Discussionmentioning

confidence: 99%

Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks

et al. 2022

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Burkholderia (B.) mallei is a host-adapted equine pathogen that causes glanders, a re-emerging zoonotic disease, which is endemic in Pakistan and other developing countries and seriously impacts the global equine movement. Due to globalization, the geographical restriction of diseases vanishes and the lack of awareness of and experience with eradicated diseases in industrialized countries also promotes the re-introduction of infections in these regions. Owing to the high equine population, the Pakistani province Punjab is a potential hotspot where several glanders outbreaks have been seen over last two decades. For determining the genomic diversity of B. mallei in this and other equine-populated prefectures, the genomes of 19 B. mallei strains isolated between 1999 and 2020 in different locations were sequenced and their genotypes were determined. Particularly, for genetically highly homogenous pathogens like B. mallei genotyping techniques require a high discriminatory power for enabling differentiation on the strain level. Thus, core-genome single nucleotide polymorphism (cgSNP) analysis was applied for distinguishing the highly similar strains. Furthermore, a whole-genome sequence-based core genome multi locus sequence typing (cgMLST) scheme, specific to B. mallei, was developed and additionally applied to the data. It was found that B. mallei genotypes in Pakistan persisted over time and space and genotype clusters preferred connection with a time point rather than the place of isolation, probably due to frequent equine movement, which promotes the spread of glanders. The cgMLST approach proved to work in accord with SNP typing and may help to investigate future glanders outbreaks.

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Section: Discussionmentioning

confidence: 99%

Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks

et al. 2022

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“…Burkholderia mallei is a small rod-shaped, Gram-negative bacterium that causes glanders, a notifiable disease in animals and humans [ 1 ]. Glanders is extremely contagious among Equidae , especially for horses and donkeys [ 2 , 3 ]. In many cases, infected animals need to be euthanized following governmental regulations on the eradication of epizootic diseases.…”

Section: Introductionmentioning

confidence: 99%

Genetic diversity and spatial distribution of Burkholderia mallei by core genome-based multilocus sequence typing analysis

et al. 2022

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Burkholderia mallei is the etiological agent of glanders, a highly contagious and often fatal disease in equids. Due to the high genetic clonality of B. mallei, high-resolution typing assays are necessary to differentiate between individual strains. Here we report on the development and validation of a robust and reproducible core genome-based Multi Locus Sequence Typing Assay (cgMLST) for B. mallei, which is based on 3328 gene targets and enables high-resolution typing at the strain level. The assay was validated using a set of 120 B. mallei genomes from public databases and 23 newly sequenced outbreak strains from in-house strain collections. In this cgMLST analysis, strains from different geographic regions were clearly distinguished by at least 70 allele differences, allowing spatial clustering while closely related and epidemiologically related strains were separated by only zero to three alleles. Neither the different sequencing technologies nor the assembly strategies had an influence on the cgMLST results. The developed cgMLST is highly robust, reproducible and can be used for outbreak investigations, source tracking and molecular characterization of new B. mallei isolates.

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“…The cELISA was further evaluated with horse sera from glanders outbreaks in Bahrain [ 9 – 10 ] and Kuwait [ 11 ]. The outbreak in Bahrain was a result from horses imported from Syria via Kuwait.…”

Section: Discussionmentioning

confidence: 99%

“…The 31 seropositive sera were obtained from two separate glanders outbreaks in the Middle East. These included 25 horse sera from a glanders outbreak in Bahrain in year 2010–2011 [ 9 – 10 ] and 6 horse sera from a glanders outbreak in Kuwait in year 2019 [ 11 ]. All serum samples were confirmed as seronegative or seropositive for glanders using the complement fixation test (CFT), which served as the gold standard in this study [ 12 ].…”

Section: Methodsmentioning

confidence: 99%

Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent

et al. 2021

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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.

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A genetic variant of Burkholderia mallei detected in Kuwait: Consequences for the PCR diagnosis of glanders

Cited by 16 publications

References 25 publications

Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks

Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks

Genetic diversity and spatial distribution of Burkholderia mallei by core genome-based multilocus sequence typing analysis

Development of a sensitive competitive enzyme-linked immunosorbent assay for serodiagnosis of Burkholderia mallei, a Tier 1 select agent

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