Performance of IS6110-LAMP assay for detection of Mycobacterium tuberculosis complex in blood and urine samples from patients with extrapulmonary tuberculosis - 01/12/23
, Rosana de Albuquerque Montenegro a, Aline dos Santos Peixoto a, Letícia Leoncio de Souza Silva b, Rayssa Maria Pastick Jares da Costa a, Wlisses Henrique Veloso Carvalho-Silva a, Danyele Costa de Mello a, George Tadeu Nunes Diniz a, Maria Almerice Lopes da Silva a, Fábio Lopes de Melo a, Lílian Maria Lapa Montenegro a, Haiana Charifker Schindler aAbstract |
Background |
The diagnosis of extrapulmonary tuberculosis (EPTB) shows numerous difficulties because of non-specific symptomatology and low sensitivity of conventional methods. Loop-mediated isothermal amplification (LAMP) is a fast and low-cost technique, which can amplify under isothermal conditions an amount of target DNA copies into approximately a billion copies.
Objective |
The present study aimed to evaluate a IS6110-LAMP system for Mycobacterium tuberculosis detection in blood and urine samples from patients with EPTB.
Methods |
The collected samples (n = 122) were stratified in two groups: Group EPTB – patient samples with confirmed EPTB (n = 61); Group non-TB – patient samples without TB (n = 61). The urine samples underwent decontamination, and the components of blood samples were separated (plasma and PBMC). DNA extractions were performed in all biological samples followed by IS6110-LAMP assay technique. The detection limit was evaluated through dilution curves (1:10) using Mtb reference strain (H37Rv) genomic DNA.
Findings |
The detection limit of IS6110-LAMP was 10 fg/μL (∼10–20 bacilli/μL). The IS6110-LAMP technique sensitivity and specificity were 95.65 % and 79.25 %, respectively, with a general kappa agreement index of 0.762.
Main conclusions |
Based on these results, IS6110-LAMP test showed considerable diagnostic parameters, being able to aid in the speed and accuracy of the final EPTB diagnosis.
Le texte complet de cet article est disponible en PDF.Highlights |
• | The IS6110-LAMP assay showed a limit detection of 2 bacili/μL. |
• | The IS6110-LAMP assay presented a high sensitivity (94.85 %). |
• | Combining IS6110-LAMP assay with other methods improved the accuracy of the EPTB diagnosis to 99.4 %. |
• | The IS6110-LAMP can aid the speed and accuracy of the final EPTB diagnosis. |
Keywords : Loop-mediated isothermal amplification, Molecular test, Tuberculosis diagnosis, Mtb, EPTB
Plan
Vol 143
Article 102423- décembre 2023 Retour au numéro
Article précédent
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