scholarly journals 2156. To Evaluate the Role of Kit-Based Loop-Mediated Isothermal Amplification (TB-LAMP) Assay in the Diagnosis of Tubercular Lymphadenitis

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S731-S732
Author(s):  
Neeraj Nischal ◽  
Saurav Das ◽  
Binit Kumar Singh ◽  
Naveet Wig ◽  
Manish Soneja ◽  
...  
Keyword(s):  
Lymph Node ◽  
Lamp Assay ◽  
Infectious Agent ◽  
Drug Susceptibility Testing ◽  
Isothermal Amplification ◽  
World Health ◽  
Reference Standard ◽  
Loop Mediated Isothermal Amplification ◽  
Tubercular Lymphadenitis ◽  
Composite Reference Standard

Abstract Background Tuberculosis (TB) is the leading cause of death as a single infectious agent worldwide. In 2017, there were an estimated 1.3 million TB deaths among HIV-negative people and 300,000 deaths among HIV-positive people. The rapid and accurate diagnosis of TB in lymphnode specimens remains a challenging task today. In 2016, World Health Organization endorsed a commercial molecular assay, the LoopAMP™ Mycobacterium tuberculosis complex (MTBC) detection kit (Eiken Chemical Company, Tokyo, Japan), which uses loop-mediated isothermal amplification (LAMP) for sputum samples only. No prospective studies on LAMP in diagnosis of Tubercular Lymphadenitis in adults have been done yet. Methods A prospective observational study with a total of 70 lymph-node aspirate specimens from suspected cases of Tubercular Lymphadenitis with age >18 years were selected and subjected to Ziehl–Neelsen staining, LAMP and culture in mycobacterial growth indicator tube (MGIT960). The immunochromatographic test was used to confirm MTB complex (MTBC) in culture positive samples and phenotypic drug susceptibility testing was done using MGIT-960. The composite reference standard (CRS) used in the study includes symptoms, radiological evidence and follow-up of 2 months. 2 × 2 tables were made and Sensitivity, Specificity, PPV, NPV of TB-LAMP were calculated with respect to AFB smear and composite reference standard (CRS). Results LAMP assay was able to detect MTBC in 34.3% (24/70) of lymph-node specimens. Sensitivity and specificity of the assay were 100% and 69.7%, respectively, considering smear as gold standard. On comparing with CRS, the assay showed 100% sensitivity and 100% specificity in the diagnosis of MTBC. Conclusion In our study, LAMP assay was found to be a promising tool for the diagnosis of Tubercular Lymphadenitis and could be used for rapid and cost-effective diagnosis of Tubercular Lymphadenitis in resource-limited settings. Disclosures All authors: No reported disclosures.

Evaluation of loop mediated isothermal amplification (LAMP) assay in the diagnosis of tubercular lymphadenitis: A pilot study

2018 ◽  
Vol 65 (1) ◽  
pp. 76-79
Author(s):  
Baijayantimala Mishra ◽  
Vinaykumar Hallur ◽  
Bijayini Behera ◽  
C. Preetam ◽  
Priti Nanda Mishra ◽  
...  
Keyword(s):  
Pilot Study ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Tubercular Lymphadenitis

scholarly journals Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Somayyeh Sedaghatjoo ◽  
Monika K. Forster ◽  
Ludwig Niessen ◽  
Petr Karlovsky ◽  
Berta Killermann ◽  
...  
Keyword(s):  
Test Performance ◽  
Genomic Dna ◽  
Lamp Assay ◽  
Fungal Species ◽  
Isothermal Amplification ◽  
Genome Comparison ◽  
Performance Study ◽  
Loop Mediated Isothermal Amplification ◽  
Tilletia Controversa ◽  
Genomic Regions

AbstractTilletia controversa causing dwarf bunt of wheat is a quarantine pathogen in several countries. Therefore, its specific detection is of great phytosanitary importance. Genomic regions routinely used for phylogenetic inferences lack suitable polymorphisms for the development of species-specific markers. We therefore compared 21 genomes of six Tilletia species to identify DNA regions that were unique and conserved in all T. controversa isolates and had no or limited homology to other Tilletia species. A loop-mediated isothermal amplification (LAMP) assay for T. controversa was developed based on one of these DNA regions. The specificity of the assay was verified using 223 fungal samples comprising 43 fungal species including 11 Tilletia species, in particular 39 specimens of T. controversa, 92 of T. caries and 40 of T. laevis, respectively. The assay specifically amplified genomic DNA of T. controversa from pure cultures and teliospores. Only Tilletia trabutii generated false positive signals. The detection limit of the LAMP assay was 5 pg of genomic DNA per reaction. A test performance study that included five laboratories in Germany resulted in 100% sensitivity and 97.7% specificity of the assay. Genomic regions, specific to common bunt (Tilletia caries and Tilletia laevis together) are also provided.

scholarly journals Development and validation of a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of ZIKV in patient samples from Brazil

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Severino Jefferson Ribeiro da Silva ◽  
Keith Pardee ◽  
Udeni B. R. Balasuriya ◽  
Lindomar Pena
Keyword(s):  
Reverse Transcription ◽  
Rapid Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Extraction ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Serum Samples ◽  
Low Resource ◽  
Loop Mediated Isothermal Amplification ◽  
One Step

AbstractWe have previously developed and validated a one-step assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of the Zika virus (ZIKV) from mosquito samples. Patient diagnosis of ZIKV is currently carried out in centralized laboratories using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR), which, while the gold standard molecular method, has several drawbacks for use in remote and low-resource settings, such as high cost and the need of specialized equipment. Point-of-care (POC) diagnostic platforms have the potential to overcome these limitations, especially in low-resource countries where ZIKV is endemic. With this in mind, here we optimized and validated our RT-LAMP assay for rapid detection of ZIKV from patient samples. We found that the assay detected ZIKV from diverse sample types (serum, urine, saliva, and semen) in as little as 20 min, without RNA extraction. The RT-LAMP assay was highly specific and up to 100 times more sensitive than RT-qPCR. We then validated the assay using 100 patient serum samples collected from suspected cases of arbovirus infection in the state of Pernambuco, which was at the epicenter of the last Zika epidemic. Analysis of the results, in comparison to RT-qPCR, found that the ZIKV RT-LAMP assay provided sensitivity of 100%, specificity of 93.75%, and an overall accuracy of 95.00%. Taken together, the RT-LAMP assay provides a straightforward and inexpensive alternative for the diagnosis of ZIKV from patients and has the potential to increase diagnostic capacity in ZIKV-affected areas, particularly in low and middle-income countries.

scholarly journals Development of a loop-mediated isothermal amplification (LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass

3 Biotech ◽  
2021 ◽  
Vol 11 (2) ◽  
Author(s):  
Domenico Rizzo ◽  
Nicola Luchi ◽  
Daniele Da Lio ◽  
Linda Bartolini ◽  
Francesco Nugnes ◽  
...  
Keyword(s):  
Lamp Assay ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Diagnostic Tools ◽  
Loop Mediated Isothermal Amplification ◽  
Larval Stages ◽  
Longhorn Beetle ◽  
Rapid Monitoring ◽  
Major Pest ◽  
Wood Borer

AbstractThe red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, where it is a major pest of cultivated and ornamental species of the genus Prunus. Morphological or molecular discrimination of adults or larval specimens is required to identify this invasive wood borer. However, recovering larval stages of the pest from trunks and branches causes extensive damage to plants and is timewasting. An alternative approach consists in applying non-invasive molecular diagnostic tools to biological traces (i.e., fecal pellets, frass). In this way, infestations in host plants can be detected without destructive methods. This paper presents a protocol based on both real-time and visual loop-mediated isothermal amplification (LAMP), using DNA of A. bungii extracted from fecal particles in larval frass. Laboratory validations demonstrated the robustness of the protocols adopted and their reliability was confirmed performing an inter-lab blind panel. The LAMP assay and the qPCR SYBR Green method using the F3/B3 LAMP external primers were equally sensitive, and both were more sensitive than the conventional PCR (sensitivity > 103 to the same starting matrix). The visual LAMP protocol, due to the relatively easy performance of the method, could be a useful tool to apply in rapid monitoring of A. bungii and in the management of its outbreaks.

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