Loop Mediated Isothermal Amplification (LAMP) Test- a Novel Nucleic Acid Based Assay for Disease Diagnosis

Rekha, V. Bhanu; Rana, Rajneesh; Arun, T.R.; Aswathi, P. B.; John, Jeny K.; Gopinath, D. M.; Sadanandan, G. V.; Jacob, Anu

doi:10.14737/journal.aavs/2014/2.6.344.350

Cited by 18 publications

(12 citation statements)

References 80 publications

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“…The amplified products can be detected by indirect detection methods like turbidity and non-specific dyes [53]. Compared to conventional PCR, LAMP assays are highly sensitive, rapid, cost-effective, and can be easily implemented in resource-limited settings [55]. However, complicated primer design and the possibility of carry-over contamination still remain a concern for LAMP-based assays [50,56].…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Amplification chemistries in clinical virology

Dunbar

Das

2019

Journal of Clinical Virology

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Molecular diagnostic methods have evolved and matured considerably over the last several decades and are constantly being evaluated and adopted by clinical laboratories for the identification of infectious pathogens. Advancement in other technologies such as fluorescence, electronics, instrumentation, automation, and sensors have made the overall diagnostic process more accurate, sensitive, and rapid. Nucleic acid based detection procedures, which rely on the fundamental principles of DNA replication have emerged as a popular and standard diagnostic method, and several commercial assays are currently available based on different nucleic acid amplification techniques. This review focuses on the major amplification chemistries that are used for developing commercial assays and discusses their application in the clinical virology laboratory. Polymerase chain reaction (PCR)The development of PCR by Kary Mullis in the 1980s revolutionized the detection of microbial pathogens in clinical specimens. PCR is a highly sensitive method that involves cyclical heating and cooling of the reaction mixture to facilitate amplification of the target DNA [3]. The method consists of three basic steps (denaturation, annealing, and extension) and the reaction mixture requires four primary components (template DNA, primers, nucleotides, and a DNA polymerase) (Fig. 1a). Double-stranded target DNA (dsDNA) is thermally denatured to form a single-stranded DNA template (ssDNA). Oligonucleotide primers anneal to complementary target sequences on the nucleic acid template and are extended by the DNA polymerase, thus creating the resultant PCR product (amplicon). The reaction occurs in a programmable thermal https://doi.

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Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Amplification chemistries in clinical virology

Dunbar

Das

2019

Journal of Clinical Virology

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“…Different techniques like Restriction Fragment Length Polymerase (RFLP) assay (Kumar et al, 2011b), Lateral Flow assay (Arun et al, 2014), Loop mediated Isothermal Amplification (LAMP) test (Rekha et al, 2014;Rekha et al, 2015) have been tried. Combination of conventional and advanced molecular tests is preferred for confirmation of field outbreaks.…”

Section: Diagnosismentioning

confidence: 99%

“…This test provides a rapid and sensitive diagnosis in a rural environment, where no laboratory equipment was available. LAMP can be performed where thermocyclers cannot be easily maintained/ procured (Rekha et al, 2014). The advantage of these tests is no need of agarose gel electrophoresis.…”

Section: Diagnosismentioning

confidence: 99%

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Mycoplasmosis in Small Ruminants in India: A Review

Yatoo¹,

Kashmir²,

Parray³

et al. 2018

JEBAS

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Mycoplasmosis, the diseases caused by mycoplasma, are one of the challenging and continuous threats to small ruminant farming. They cause heavy morbidity (upto 100%), mortality (10-100%) and huge economic loss. Common diseases caused by mycoplasma organisms are contagious caprine pleuropneumonia (mostly in goats), contagious agalactia (both sheep and goats), atypical pneumonia (usually sheep), besides arthritis, mastitis, seminal vesiculitis, ampullitis, epididymitis, orchitis, urethritis, conjunctivitis and meningitis either alone or as classical syndromes. The common mycoplasma includes Mycoplasma mycoides subspecies mycoides, M. mycoides subsp. capri, M. capricolum subsp. capripneumoniae, M. capricolum subsp. capricolum, M. agalactiae, M. bovis and M. conjunctivae etc. Most of the mycoplasmas produce a conglomerate of clinical signs hampering diagnosis. The other main challenge posed by mycoplasma is the difficulty in isolation due to lack of

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“…(such as a thermocycler) and post amplification processing and are still time-consuming to accomplish at chair-side [3]; these drawbacks led to the emergence of the alternative isothermal amplification methods which are easier and not requiring thermal cycling as these methods amplify nucleic acids to large copies under isothermal conditions [4]. Several isothermal amplification protocols have been generated over the last 10 years, one of which loop-mediated isothermal amplification (LAMP).…”

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confidence: 99%

Colorimetric LAMP Molecular Method for Immediate Detection of Three Periodontal Pathogens

Al-Hamdoni¹,

Al-Rawi²

2020

Preprint

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Background and Purpose: For enhancing the efficiency of diagnostic tools and overcome drawbacks faced in diagnosis attempt, molecular methods have been developed aid accurate and rapid identification of bacterial species particularly those which are difficult to study with. The current study searched to find a more specific and rapid method serves researchers for the identification of periodontal pathogens and overcome disturbances encountered in study such anaerobic fastidious organisms. Methods and Results: Three periodontal pathogens, Porphyromonas gingivalis , Tannerella forsythia and Treponema denticola were characterized and identified by phenotypic features and Loop- Mediated Isothermal Amplification (LAMP) techniques which involved 4 sets of primers targeted to 16SrRNA genes and loop primers and the Colorimetric Master Mix containing Bst DNA polymerase and Phenol Red for the detection of amplicon formation. There was a variance in phenotypic characters of the isolates of the same species. LAMP, as a novel molecular technique was of usefulness value in identifying the target weather as extracted DNA or whole cells in a high specific and very rapid manner within 30 min. by visual reading of the results. Conclusion: It is strongly recommended the use of the novel LAMP method in researches work with periodontal pathogens as its advantages make it superior to other molecular techniques in respects of a higher specificity, rapidity, sensitivity and overcome DNA extraction step and special equipment in that it can be used at chair- side identification.

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Loop Mediated Isothermal Amplification (LAMP) Test- a Novel Nucleic Acid Based Assay for Disease Diagnosis

Cited by 18 publications

References 80 publications

Amplification chemistries in clinical virology

Amplification chemistries in clinical virology

Mycoplasmosis in Small Ruminants in India: A Review

Colorimetric LAMP Molecular Method for Immediate Detection of Three Periodontal Pathogens

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