A recombinase polymerase amplification lateral flow assay for rapid detection of Burkholderia pseudomallei, the causative agent of melioidosis

2022 ◽  
Author(s):  
Apoorva Saxena ◽  
Vijai Pal ◽  
Nagesh Kumar Tripathi ◽  
Ajay Kumar Goel
Keyword(s):  
Causative Agent ◽  
Rapid Detection ◽  
Burkholderia Pseudomallei ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Recombinase Polymerase Amplification

scholarly journals Rapid Detection of Burkholderia pseudomallei with a Lateral Flow Recombinase Polymerase Amplification Assay

2019 ◽  
Author(s):  
Yao Peng ◽  
Zheng Xiao ◽  
Biao Kan ◽  
Wei Li ◽  
Wen Zhang ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Burkholderia Pseudomallei ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Spiked Soil ◽  
Lateral Flow Dipstick ◽  
Life Threatening ◽  
Amplification Assay

AbstractMelioidosis is a severe infectious disease caused by gram-negative, facultative intracellular pathogen Burkholderia pseudomallei (B. pseudomallei). Although cases are increasing reported from other parts of the world, it is an illness of tropical and subtropical climates primarily found in southeast Asia and northern Australia. Because of a 40% mortality rate, this life-threatening disease poses a public health risk in endemic area. Early detection of B. pseudomallei infection benefits greatly to implement effective treatment timely, which is vital for prognosis of a melioidosis patient. In this study, a novel isothermal recombinase polymerase amplification combined with lateral flow dipstick (LF-RPA) assay was established for rapid detection of B.pseudomallei. A set of probe and primers targeting orf2 gene of B. pseudomallei were generated and parameters for the LF-RPA assay were optimized. Result can be easy visualized in 30 minutes with the limit of detection (LoD) as low as 20 femtogram (ca. 25.6 copies) of B. pseudomallei genomic DNA. The assay is highly specific as no cross amplification was observed with 35 non-B. pseudomallei pathogens. Isolates (N=19) from patients of Hainan province of China were retrospectively confirmed by the newly developed method. LoD for B. pseudomallei spiked soil and blood samples were 2.1×103 CFU/g and 4.2×103 CFU/ml respectively. Sensitivity of the LF-RPA assay was comparable to TaqMan Real-Time PCR, however, the LF-RPA assay exhibited a better tolerant to inhibitors in blood than the later. Our results showed that the LF-RPA assay is an alternative to existing PCR-based methods for detection of B. pseudomallei with a potentiality of early accurate diagnosis of melioidosis at point of care or in-field use.

Development of a PCR-lateral flow assay for rapid detection of Yersinia pestis, the causative agent of plague

Acta Tropica ◽  
2021 ◽  
pp. 105958
Author(s):  
Rita Singh ◽  
Vijai Pal ◽  
Manoj Kumar ◽  
N.K Tripathi ◽  
A.K. Goel
Keyword(s):  
Yersinia Pestis ◽  
Causative Agent ◽  
Rapid Detection ◽  
Lateral Flow ◽  
Lateral Flow Assay
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