Development of probe‐based real‐time loop‐mediated isothermal amplification for detection of<i>Brucella</i>

Bhat, Irfan Ahmad; Mashooq, Mohmad; Kumar, Dinesh; Varshney, Rajat; Rathore, Rajesh

doi:10.1111/jam.13938

Cited by 13 publications

(9 citation statements)

References 15 publications

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“…Molecular biological detection of Brucella is abundant [9,20,21]. However, there were a few PCR methods for detecting canine Brucella [10,11].…”

Section: Discussionmentioning

confidence: 99%

“…Its effectiveness is affected by the infected animal's bacteremia level [7]. Molecular biological detection technology has the characteristics of safety and reliability, high sensitivity, strong speci city and simple operation, and has begun to be popularized and applied to detect Brucella [8,9]. Now there were a variety of PCR methods for detecting canine Brucella [10,11], such as bruce-ladder [11] and multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) procedures [12].…”

Section: Introductionmentioning

confidence: 99%

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A New Specific Sequence to Distinguish B.canis From Other Brucella by PCR

Ye¹,

Yang²,

Li³

et al. 2021

Preprint

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Background: Brucellosis is a zoonotic disease worldwide. The increasing number of pet dogs has raised new risk of people getting canine brucella with absent or mild symptom. Besides, the canine brucellosis can be caused by other brucella species, so their infection could be omitted by the PCR method. The present PCR methods can only detect canine brucella, by which cases infected with other brucella would appear negative. It’s an urge to develop a specific PCR assay for detecting canine Brucellosis, Whether the pathogen was B.canis or any other Brucella.Resaults: a differential sequence of B.canis were found by genome comparison analysis and were analyzed by BLAST. Then a PCR method was established using specific primers in the sequence and tested for clinical application. It could detect canine brucellosis caused by B.canis or other Brucella species with 310-bp and 413-bp product, respectively. The developed PCR method had specificity for non-brucella and a sensitivity of 100 copies of Brucella DNA. The detection accuracy verified with spiked samples was 95.5% (21/22) for B.canis and 100% (22/22) for other brucella. Conclusions: The study found a specific sequence of B.canis and developed a PCR detection method to detect canine brucellosis caused by B.canis or other Brucella species. The method established in this study will more comprehensively detect the pathogen of canine brucellosis and provide important methods and means for preventing and controlling this disease.

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“…Molecular biological detection of Brucella is abundant [9,20,21]. However, there were a few PCR methods for detecting canine Brucella [10,11].…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A New Specific Sequence to Distinguish B.canis From Other Brucella by PCR

Ye¹,

Yang²,

Li³

et al. 2021

Preprint

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“…27,68 DARQ LAMP was also applied to detect Salmonella, 69 avian reovirus, 70 methicillin-resistant Staphylococcus aureus (MRSA) 71 and Brucella. 72 Analytical sensitivity for Salmonella DARQ LAMP was 10 copies per reaction. High specicity against non-Salmonella strains was shown.…”

Section: Alternately Binding Quenching Probe Competitive Lamp (Abc-lamp)mentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP) – review and classification of methods for sequence-specific detection

Becherer

Borst

Bakheit³

et al. 2020

Anal. Methods

289

245

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“…In this study, we developed a multiplex SFTSV/OT/IC loop‑mediated isothermal amplification (LAMP) assay for the differential diagnosis of SFTSV and O . tsutsugamushi infection, based on a slightly modified DARQ probe method [ 33 , 34 ]. The detection limit of the multiplex SFTSV/OT/IC RT-LAMP assay was confirmed using diluted SFTS and scrub typhus clinical samples.…”

Section: Introductionmentioning

confidence: 99%

Developing a multiplex loop-mediated isothermal amplification assay (LAMP) to determine severe fever with thrombocytopenia syndrome (SFTS) and scrub typhus

et al. 2022

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Severe fever with thrombocytopenia syndrome (SFTS) and scrub typhus are endemic zoonotic diseases that pose significant public health threats in East Asia. As these two diseases share common clinical features, as well as overlapping disease regions, it is difficult to differentiate between SFTS and scrub typhus. A multiplex reverse-transcription loop‑mediated isothermal amplification (RT-LAMP) assay was developed to detect large segments and GroES genes for SFTS virus (SFTSV) and Orientia tsutsugamushi (OT). The performance of the RT-LAMP assay was compared and evaluated with those of commercial PowerChek™ SFTSV real-time PCR and LiliF™ TSUTSU nested PCR for 23 SFTS and 12 scrub typhus clinical samples, respectively. The multiplex SFTSV/OT/Internal control (IC) RT-LAMP assay showed comparable sensitivity (91.3%) with that of commercial PowerChek™ SFTSV Real-time PCR (95.6%) and higher sensitivity (91.6%) than that of LiliF™ TSUTSU nested PCR (75%). In addition, the multiplex SFTSV/OT RT-LAMP assay showed 100% specificity and no cross-reactivity for blood from uninfected healthy patients and samples from patients infected with other fever viruses. Thus, the multiplex SFTSV/OT/IC RT-LAMP assay could serve as a useful point-of-care molecular diagnostic test for SFTS and scrub typhus.

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Development of probe‐based real‐time loop‐mediated isothermal amplification for detection ofBrucella

Abstract: The developed probe based RT-LAMP is a powerful gene amplification technique is a specific, fast diagnostic tool for early detection and identification of Brucella. This article is protected by copyright. All rights reserved.

Cited by 13 publications

References 15 publications

A New Specific Sequence to Distinguish B.canis From Other Brucella by PCR

A New Specific Sequence to Distinguish B.canis From Other Brucella by PCR

Loop-mediated isothermal amplification (LAMP) – review and classification of methods for sequence-specific detection

Developing a multiplex loop-mediated isothermal amplification assay (LAMP) to determine severe fever with thrombocytopenia syndrome (SFTS) and scrub typhus

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