Feasibility and Operational Performance of Tuberculosis Detection by Loop-Mediated Isothermal Amplification Platform in Decentralized Settings: Results from a Multicenter Study

Abstract

Currently available nucleic acid amplification platforms for tuberculosis (TB) detection are not designed to be simple or inexpensive enough to implement in decentralized settings in countries with a high burden of disease. The loop-mediated isothermal amplification platform (LAMP) may change this. We conducted a study in adults with symptoms suggestive of TB in India, Uganda, and Peru to establish the feasibility of using TB-LAMP (Eiken Chemical Co.) in microscopy laboratories compared with using smear microscopy against a reference standard of solid and liquid cultures. Operational characteristics were evaluated as well. A total of 1,777 participants met the eligibility criteria and were included for analysis. A pproximately 9 million people developed tuberculosis (TB), and 1.5 million people died from the disease in 2013 (1). Substantial progress has been made in increasing treatment success, but 2.9 million of these cases were not diagnosed or diagnosis was not reported due to limited access to diagnostic tests, insufficient sensitivity, or long turnaround time. More rapid and accurate detection at lower levels of care is necessary to reach these lost and missed cases (2). Nucleic acid amplification tests (NAATs) offer speed and sensitivity for pathogen detection, but until recently, no commercial systems had been designed to be simple or inexpensive enough for decentralized settings (3).Xpert MTB/RIF (Cepheid Inc., Sunnyvale, CA), a rapid automated molecular test, has been a major step in the right direction, but initial investment in the equipment and subsequent maintenance remain quite expensive. The development of a less costly approach through the use of manual techniques may expand the application of NAATs.TB-LAMP is a manual TB detection method that is based on the novel loop-mediated isothermal amplification platform (LAMP) from Eiken Chemical Co. in Japan. LAMP has several features that make it attractive as a diagnostics platform for resource-poor settings. It takes less than 2 h (Ͻ60 min of hands-on time), it is less complex than traditional molecular methods, and it generates a fluorescent result that can be detected with the naked eye (4). In a recent study in peripheral laboratories in China using the TB-LAMP assay, the sensitivities in smear-positive, culturepositive sputum specimens and smear-negative, culture-positive sputum specimens were 92.1% (152/165; 95% confidence interval [CI], 86.9% to 95.3%) and 53.8% (113/210; 95% CI, 47.1% to 60.4%), respectively, using solid culture as a reference. The specificity in culture-negative samples was 98.3% (938/954; 95% CI, 97.3% to 99.0%) (5).In this study, we aimed to evaluate the feasibility and operational characteristics of LAMP in microscopy centers, including Citation Gray CM, Katamba A, Narang P, Giraldo J, Zamudio C, Joloba M, Narang R, Paramasivan CN, Hillemann D, Nabeta P, Amisano D, Alland D, Cobelens F, Boehme CC. 2016. Feasibility and operational performance of tuberculosis detection by loop-mediated isothermal amplification platform in ...

Establishment of an accurate and fast detection method using molecular beacons in loop-mediated isothermal amplification assay

“…Although the recognition of 6–8 independent target sequences (via 4–6 primers) suggests higher specificity and sensitivity than standard PCR, which uses only two primers, the increase in primers can also cause nonspecific amplification9. Compounding this problem is the fact that traditional methods of evaluating LAMP reactions cannot directly check for nonspecific amplification, among other issues.…”

Clinical Performances of Pure TB-Lamp Kit for <i>M. tuberculosis</i> Complex Detection in Sputum Samples

“…This study shows that for active tuberculosis detection in routine, Sensitivity of TB-Lamp assay is higher than SSM. Previous studies conducted in differents setting have demonstrated that TB-Lamp assay has an excellent sensitivity for Tuberculose detection [10]- [15].…”

“…This method is dependent on autocycling strand displacement DNA synthesis performed by the Bst DNA polymerase large fragment which has high-strand displacement activity, and is carried out at 65˚C for 1 hour [9]. TB-Lamp assay seems well adapted for in low incomes countries [6] [10].…”

Diagnosis of Tuberculosis: Nanodiagnostics Approaches

“…The unique property depends on the size and composition of the particles compared to atoms and other materials. These properties includes: (1) large to volume ratio (metal NP, in The sensitivity of TB-LAMP is lower than Xpert MTB/RIF assay but greater than smear microscopy (Gray et al 2016;Pham et al 2018) Recommended by WHO particular gold NP), (2) surface plasmon resonance, (3) Surface-Enhanced Raman Scattering (SERS), (4) super-magnetization or ferromagnetic nanoparticles (e.g. iron oxide), (5) enhanced photoluminescence (semiconductor quantum dots), (6) high electric and heat conductivity, (7) potent surface catalytic activity (Gatoo et al 2014;Khan et al 2019).…”