Development of the Novel Loop Mediated Isothermal Amplification (LAMP) of IS711 Sequence for Rapid Detection of Brucella Species

Zadon, Stanzin; Sharma, N S; Arora, Anil; Mudit, Chandra

doi:10.1007/s40011-014-0377-9

Cited by 2 publications

(4 citation statements)

References 25 publications

(29 reference statements)

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“…Brucellosis, which is one of the worldwide zoonoses, is still of public health, veterinarian, and economic concern in many regions of the world 1. Brucellosis is caused by a number of host adapted species of facultative intracellular bacteria of the genus Brucella 2…”

Section: Introductionmentioning

confidence: 99%

“…To overcome the drawbacks posed by PCR-based assays, various other techniques, on the basis of isothermal amplification of nucleic acid, have been designed, which are able to rapidly synthesize DNA in large amounts without the use of any sophisticated apparatus 11. Among these isothermal amplification methods, loop-mediated isothermal amplification (LAMP) is one of the most widely used techniques, which has been employed for rapid, sensitive, and specific detection of many bacterial pathogens including Brucella spp 1,12–16. However, the results of Brucella -LAMP assays were displayed using agarose gel electrophoresis, color indicator (such as hydroxynaphthol blue, SYBR green I, calcein dye et al), and real-time turbidity equipment.…”

Section: Introductionmentioning

confidence: 99%

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Lateral flow biosensor combined with loop-mediated isothermal amplification for simple, rapid, sensitive, and reliable detection of Brucella spp

Liu

Wang

et al. 2019

IDR

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Brucella species is responsible for brucellosis in human and animals, which is still of public health, veterinarian, and economic concern in many regions of the world. Here, a novel molecular diagnosis assay, termed loop-mediated isothermal amplification coupled with nanoparticles-based lateral flow biosensor (LAMP-LFB), was developed and validated for simply, rapidly, and reliably detecting all Brucella spp. strains. A set of six primers was designed based on the Brucella -specific gene Bscp31 . The Brucella -LAMP results were visually reported by biosensor within 2 mins. A variety of bacterial strains representing several Brucella species, as well as several Gram-negative and Gram-positive bacterial species were used to determine the analytical sensitivity and specificity of the assay. Optimal LAMP conditions were 63°C for 40 mins, and the assay’s sensitivity was found to be 100 fg of genomic DNA in the pure cultures. No cross-reactions to non- Brucella strains were obtained; thus, analytical specificity of LAMP-LFB assay is of 100%. Using the protocol, 20 mins for rapid DNA preparation followed by isothermal amplification (40 mins) combined with biosensor detection (2 mins) resulted in a total assay time of approximately 65 mins. In the case of 117 whole blood samples, 13 (11.11%) samples were Brucella -positive by LAMP-LFB, and the diagnostic accuracy was 100% when compared to the culture-biotechnical method. In conclusion, Brucella -LAMP-LFB technique developed in this study is a sensitive and specific method to rapidly identify all Brucella spp. strains, and can be applied as a potential diagnostic tool for brucellosis in basic, clinical, and field laboratories.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Lateral flow biosensor combined with loop-mediated isothermal amplification for simple, rapid, sensitive, and reliable detection of Brucella spp

Liu

Wang

et al. 2019

IDR

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“…Certain potential targets have been reported for diagnosis of brucellosis through LAMP technique viz. bcsp31 (Ohtsuki et al, 2008), omp25 (Lin et al, 2011;Pan et al, 2011;Song et al, 2012;Chen et al, 2013;Soleimani et al, 2013) and IS711 (Perez-Sancho et al, 2013;Zadon et al, 2014). Here we have reported a novel potential target, i.e.…”

Section: Introductionmentioning

confidence: 53%

“…Further, omp25-LAMP was reported to be 10 times more sensitive than PCR (Song et al, 2012;Karthik et al, 2014). IS711-LAMP was found 100 times more sensitive than conventional PCR (Zadon et al, 2014…”

Section: Analytical Specificity and Relative Sensitivitymentioning

confidence: 99%

A Novel Close-tube Loop Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Brucella

Mishra

Thomas

Jeena

et al. 2021

IJAR

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Background: Brucellosis is one of the important diseases affecting both human as well as livestock. Rapid diagnosis of the pathogen is highly essential for undertaking effective therapeutic measures. Loop mediated isothermal amplification (LAMP) assay, one of the robust nucleic acid detection platforms, is used especially for rapid disease diagnosis. However, persistent contamination has been the major bottleneck in LAMP which can be eliminated with adoption of appropriate closed-tube formats.Methods: In the present report, two sets of LAMP primers targeting omp2b gene of Brucella were designed and standardized for detection of all the major Brucella spp. Result: There was absence of amplification in any of the non-Brucella species in contrast to detection of white precipitation in unaided eye in daylight as well as greenish fluorescence under ultraviolet light in all of the Brucella species. It was found to be more sensitive than conventional PCR as relative sensitivity was found to be 0.34pg for first set of primers and 34fg for second set of primers, as compared to 3.4pg through bcsp31 PCR.

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Development of the Novel Loop Mediated Isothermal Amplification (LAMP) of IS711 Sequence for Rapid Detection of Brucella Species

Cited by 2 publications

References 25 publications

<p>Lateral flow biosensor combined with loop-mediated isothermal amplification for simple, rapid, sensitive, and reliable detection of <em>Brucella</em> spp</p>

<p>Lateral flow biosensor combined with loop-mediated isothermal amplification for simple, rapid, sensitive, and reliable detection of <em>Brucella</em> spp</p>

A Novel Close-tube Loop Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Brucella

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