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Development of a rapid and sensitive recombinase polymerase amplification-lateral flow assay for detection of Burkholderia mallei.
Abstract: Burkholderia mallei, a potential biothreat agent is the aetiological agent of glanders, a zoonotic disease primarily affecting equines. B. mallei shares close genetic proximity with B. pseudomallei, the aetiological agent of melioidosis. Hence, molecular detection of B. mallei and its differentiation from B. pseudomallei has always been challenging. Early diagnosis of glanders is critical for timely treatment in humans and disease containment in animals. In this study a recombinase polymerase amplification-lateral flow (RPA-LF) assay has been developed for early and accurate detection of B. mallei. RPA-LF assay was found to be highly sensitive and detected as low as 10 fg genomic DNA of B. mallei. The assay was highly specific and could differentiate B. mallei and B. pseudomallei. The assay also detected B. mallei in artificially spiked blood, tap water and garden soil. The established assay is simple, rapid and does not require complex instrumentation. The field deployable assay can have better implications in rapid glanders diagnosis and environmental detection of B. mallei over PCR-based detection tools in glanders endemic areas with limited laboratory resources.
© 2019 Blackwell Verlag GmbH.
Publication Date: 2019-02-15 PubMed ID: 30650261DOI: 10.1111/tbed.13126Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Burkholderia mallei
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Management
- Disease Outbreaks
- Disease Prevalence
- Disease Surveillance
- Disease Transmission
- Disease Treatment
- Epidemiology
- Equine Health
- Glanders
- Infection
- Infectious Disease
- Molecular biology
- Public Health
- Veterinary Medicine
- Veterinary Research
- Zoonotic Diseases
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The researchers in this article have developed a quick and sensitive tool, the recombinase polymerase amplification-lateral flow (RPA-LF) assay, for detecting Burkholderia mallei, a bacterial agent responsible for causing glanders. This assay can differentiate between B. mallei and closely related species B. pseudomallei, offering a valuable tool for diagnosing of glanders and potentially preventing the spread of the disease.
Background of the Study
- Burkholderia mallei is a potent biothreat and cause of zoonotic disease glanders. The ailment mainly affects equines, but humans can also be infected. Timely detection of the bacterium is crucial for early treatment of the disease in humans and its containment in animals.
- This species has a close genetic relationship with Burkholderia pseudomallei, which causes another disease known as melioidosis. Thus, accurately detecting B. mallei and distinguishing it from B. pseudomallei is a considerable challenge.
Introduction to Recombinase Polymerase Amplification-Lateral Flow (RPA-LF) Assay
- The researchers developed a tool, known as recombinase polymerase amplification-lateral flow (RPA-LF) assay, to overcome this challenge.
- RPA-LF assay is a simple and rapid diagnostic tool that does not need sophisticated devices to operate. This makes it an ideal instrument for use in locations with limited access to high-tech laboratory resources.
Performance of the RPA-LF Assay
- The RPA-LF assay was found to be highly sensitive, detecting as little as 10fg of B. mallei’s genomic DNA.
- It also successfully differentiated between B. mallei and B. pseudomallei, proving its specificity.
- In addition to its high sensitivity and specificity, the assay also successfully detected B. mallei in experimental trials involving artificially spiked samples of blood, tap water, and garden soil.
Implications of the Study
- The RPA-LF assay is positioned to have broader implications for glanders diagnosis and environmental detection of B. mallei.
- It has the potential to replace PCR-based detection tools, especially in areas endemic to glanders where resources are limited.
Cite This Article
APA
Saxena A, Pal V, Tripathi NK, Goel AK.
(2019).
Development of a rapid and sensitive recombinase polymerase amplification-lateral flow assay for detection of Burkholderia mallei.
Transbound Emerg Dis, 66(2), 1016-1022.
https://doi.org/10.1111/tbed.13126 Publication
Researcher Affiliations
- Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior, India.
- Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior, India.
- Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior, India.
- Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior, India.
MeSH Terms
- Animals
- Blood / microbiology
- Burkholderia mallei / genetics
- Burkholderia mallei / isolation & purification
- DNA, Bacterial / genetics
- Horses
- Humans
- Nucleic Acid Amplification Techniques / methods
- Nucleic Acid Amplification Techniques / veterinary
- Recombinases
- Soil Microbiology
- Water Microbiology
Grant Funding
- DRDO
Citations
This article has been cited 10 times.- Gassiep I, Chewapreecha C, Chantratita N, Oakley T, Mukhopadhyay C, Lal PB, AuCoin D, Chowdhury FR, Meumann EM, Currie BJ, Dance DAB, Wuthiekanun V, Norton R. Laboratory diagnosis of melioidosis. PLoS Negl Trop Dis 2025 Dec;19(12):e0013761.
- Daddy Gaoh S, Kweon O, Ahn Y. Propidium Monoazide (PMAxx)-Recombinase Polymerase Amplification Exo (RPA Exo) Assay for Rapid Detection of Burkholderia cepacia Complex in Chlorhexidine Gluconate (CHX) and Benzalkonium Chloride (BZK) Solutions. Microorganisms 2023 May 26;11(6).
- Hifumi T, Tanaka T, Sato M, Akioka K, Fujimata C, Miyoshi N. Rapid detection of alveolar echinococcosis in hepatic nodules of horses by recombinase polymerase amplification assay. Vet Anim Sci 2023 Jun;20:100291.
- Saxena A, Rai P, Mehrotra S, Baby S, Singh S, Srivastava V, Priya S, Sharma SK. Development and Clinical Validation of RT-LAMP-Based Lateral-Flow Devices and Electrochemical Sensor for Detecting Multigene Targets in SARS-CoV-2. Int J Mol Sci 2022 Oct 28;23(21).
- Li J, Zhong Q, Shang MY, Li M, Jiang YS, Zou JJ, Ma SS, Huang Q, Lu WP. Preliminary Evaluation of Rapid Visual Identification of Burkholderia pseudomallei Using a Newly Developed Lateral Flow Strip-Based Recombinase Polymerase Amplification (LF-RPA) System. Front Cell Infect Microbiol 2021;11:804737.
- Saxena A, Pal V, Tripathi NK, Goel AK. A recombinase polymerase amplification lateral flow assay for rapid detection of Burkholderia pseudomallei, the causative agent of melioidosis. Braz J Microbiol 2022 Mar;53(1):185-193.
- Panpru P, Srisrattakarn A, Panthasri N, Tippayawat P, Chanawong A, Tavichakorntrakool R, Daduang J, Wonglakorn L, Lulitanond A. Rapid detection of Enterococcus and vancomycin resistance using recombinase polymerase amplification. PeerJ 2021;9:e12561.
- Deng Y, Jiang H, Li X, Lv X. Recent advances in sensitivity enhancement for lateral flow assay. Mikrochim Acta 2021 Oct 13;188(11):379.
- Dong Y, Zhao P, Chen L, Wu H, Si X, Shen X, Shen H, Qiao Y, Zhu S, Chen Q, Jia W, Dong J, Li J, Gao S. Fast, simple and highly specific molecular detection of Vibrio alginolyticus pathogenic strains using a visualized isothermal amplification method. BMC Vet Res 2020 Mar 4;16(1):76.
- Moawad AA, Silge A, Bocklitz T, Fischer K, Rösch P, Roesler U, Elschner MC, Popp J, Neubauer H. A Machine Learning-Based Raman Spectroscopic Assay for the Identification of Burkholderia mallei and Related Species. Molecules 2019 Dec 10;24(24).
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