A real-time loop mediated isothermal amplification assay for molecular detection of Burkholderia mallei, the aetiological agent of a zoonotic and re-emerging disease glanders

Saxena, Apoorva; Pal, Vijai; Tripathi, Nagesh K.; Goel, Ajay

doi:10.1016/j.actatropica.2019.04.004

Cited by 12 publications

(20 citation statements)

References 18 publications

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“…Similarly, the LAMP assay reported by Pal et al [ 31 ] could detect as low as 1 pg of B. mallei NCTC 10230. Saxena et al [ 26 ] has determined B. mallei NCTC 10245 using as little as by 250 fg of genomic DNA. Furthermore, the LAMP assay could detect 4.73 × 10 2 –2.1 × 10 3 CFU/mL B. mallei NCTC 10245 of artificially spiked tap water and human blood [ 26 ].…”

Section: Discussionmentioning

confidence: 99%

“…Saxena et al [ 26 ] has determined B. mallei NCTC 10245 using as little as by 250 fg of genomic DNA. Furthermore, the LAMP assay could detect 4.73 × 10 2 –2.1 × 10 3 CFU/mL B. mallei NCTC 10245 of artificially spiked tap water and human blood [ 26 ]. In this study, the LAMP assay using 10 CFU/mL inocula provided high detection capability, showing sensitivity of 58.3% (70/120, 70 positives out of 120 samples) in autoclaved nuclease-free water, 24.2% (29/120) in CHX, and 35% (42/120) in BZK.…”

Section: Discussionmentioning

confidence: 99%

“…Although the LAMP assay has shown to detect B. mallei and B. pseudomallei despite limitations [ 24 , 26 , 31 ], it depended on the type of various non-sterilized pharmaceutical products. Based on the simplest methods to get the shortest procedure, including DNA extraction by boiling procedure, the cost of LAMP per test is also considerably lower than other available molecular tests [ 37 ].…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, the reaction can be performed faster and at a much lower cost than by other PCR methods. Due to the high efficiency and sensitivity of LAMP, this method has been widely applied to pathogen detection [ 21 , 22 , 23 , 24 , 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting Burkholderia cepacia Complex in Non-Sterile Pharmaceutical Products

et al. 2021

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Simple and rapid detection of Burkholderia cepacia complex (BCC) bacteria, a common cause of pharmaceutical product recalls, is essential for consumer safety. In this study, we developed and evaluated a ribB-based colorimetric loop-mediated isothermal amplification (LAMP) assay for the detection of BCC in (i) nuclease-free water after 361 days, (ii) 10 μg/mL chlorhexidine gluconate (CHX) solutions, and (iii) 50 μg/mL benzalkonium chloride (BZK) solutions after 184 days. The RibB 5 primer specifically detected 20 strains of BCC but not 36 non-BCC strains. The limit of detection of the LAMP assay was 1 pg/μL for Burkholderia cenocepacia strain J2315. Comparison of LAMP with a qPCR assay using 1440 test sets showed higher sensitivity: 60.6% in nuclease-free water and 42.4% in CHX solution with LAMP vs. 51.3% and 31.1%, respectively, with qPCR. These results demonstrate the potential of the ribB-based LAMP assay for the rapid and sensitive detection of BCC in pharmaceutical manufacturing.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting Burkholderia cepacia Complex in Non-Sterile Pharmaceutical Products

et al. 2021

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“…Species of the genera (Escherichia, Salmonella, Shigella) are considered the main enteropathogens, causing diarrheal diseases in humans through food ingestion, many times coming directly from agriculture [26,29,84,85]. Similarly, Burkholderia mallei of the Burkholderiaceae family is considered pathogenic, causing "glanders" disease (contagious and re-emerging) in horses, donkeys and mules, but with low probability of transmission to humans, except for direct contacts with sick animals and other media [86]. Several genera of the Enterobacteriaceae family, such as Enterobacter, Escherichia, Klebsiella, Salmonella and Shigella, can be found in both soil and water; they include bacteria recognized as part of the microbiota of the digestive tract of animals such as cattle, horses and even humans, which can be natural dispersers to these ecosystems [87].…”

Section: Discussionmentioning

confidence: 99%

Gram-Negative Bacteria from Organic and Conventional Agriculture in the Hydrographic Basin of Loja: Quality or Pathogen Reservoir?

et al. 2021

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Organic and conventional agriculture are vital for the development of human society; however, the use of contaminated water and the inappropriate use of organic chemical fertilizers can lead to an increase in the microbial load (potentially pathogenic) of the normal microbiota of the agricultural soil. In this context, the aim of our study was to isolate Gram-negative bacteria from the superficial soil layer and irrigation water of agricultural areas (11 organic farms and nine conventional farms) and consider their potential ecological and health risk importance. Through culture isolation using three bacterial media (TSA) trypticase soy agar (general nutritive media); MacConkey Gram-negative bacteria and (EMB) eosin methylene blue agar (selective for Enterobacteriaceae) and classical biochemical tests, we recorded a total of 12 bacterial species, most belonging to the Enterobacteriaceae family, such as Enterobacter, Escherichia, Klebsiella, Salmonella and Shigella, which can be pathogenic for humans and animals. In contrast, bacteria such as Pantoea agglomerans, Pseudomonas aeruginosa, P. fluorescens and Burkholderia mallei could facultatively work as diazotrophic or plant growth-promoting rhizobacteria. Soil bacteria richness detected with the media applied was significantly higher than water bacteria, but we found no significant differences between organic and conventional agriculture. We conclude that the isolated bacteria in water and soil mostly belongs to enteropathogenic bacteria which could be pathogenic to animals and humans. While other bacteria like Pseudomonas aeruginosa could be viewed as useful by improving nutrient availability in agricultural soil.

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Standardization of loop-mediated isothermal amplification for detection of D. nodosus and F. necrophorum causing footrot in sheep and goats

Kavitha

Kumar

Sreedevi

2022

Trop Anim Health Prod

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The Loop Mediated Isothermal Ampli cation (LAMP) was standardized for rapid detection of D. nodosus and F. necrophorum. A total of 250 foot swabs (200) were screened from sheep and (50) were from goats from different districts of Rayalaseema viz., Chittoor, Nellore, Kadapa, Anantapur. Out of 250 samples 75 (30.0%) and 85 (34.0%) were positive for D. nodosus and F. necrophorum respectively. All the 250 samples were screened individually for both the organisms by LAMP. Among them, 104 (41.6%) were found to be positive for D. nodosus and 120 (48.0%) were positive for F. necrophorum. The e cacy of LAMP in terms of sample DNA detection limit was compared with the PCR by using standard dilutions of DNA extracted from D. nodosus and F. necrophorum cultures. The detection limit was found to be higher than PCR for both the organisms. The sensitivity of LAMP is compared with PCR by targeting 16S rRNA gene of D. nodosus and lktA gene of F. necrophorum. In case of D. nodosus, out of 250 samples, 75 (30.0%) were positive by PCR and 104 (41.6%) were positive by LAMP. Among 250 samples, 85 (34.0%) were positive by PCR and 120 (48.0%) were positive by LAMP in case of F. necrophorum. The LAMP was found to be more sensitive than PCR in detecting the organisms with high statistical signi cance.

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A real-time loop mediated isothermal amplification assay for molecular detection of Burkholderia mallei, the aetiological agent of a zoonotic and re-emerging disease glanders

Cited by 12 publications

References 18 publications

Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting Burkholderia cepacia Complex in Non-Sterile Pharmaceutical Products

Loop-Mediated Isothermal Amplification (LAMP) Assay for Detecting Burkholderia cepacia Complex in Non-Sterile Pharmaceutical Products

Gram-Negative Bacteria from Organic and Conventional Agriculture in the Hydrographic Basin of Loja: Quality or Pathogen Reservoir?

Standardization of loop-mediated isothermal amplification for detection of D. nodosus and F. necrophorum causing footrot in sheep and goats

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