Original articleRapid and effective diagnosis of pulmonary tuberculosis with novel and sensitive loop-mediated isothermal amplification (LAMP) assay in clinical samples: A meta-analysis
Introduction
Tuberculosis (TB) remains a serious health problem worldwide with nearly 9 million new cases and 1.4 million deaths annually [1]. Several publications and international guidelines highlight the need for appropriate and accurate diagnosis for effective treatment. In this regard, the development of novel and accurate diagnostic tools which may supplement or replace existing methods becomes a necessity.
Rapid and effective diagnosis of patients suspected of having TB remains a challenge. The conventional TB diagnosis techniques, including methods based on direct microscopic examination by ZiehleNeelsen staining, culture, chest radiography and tuberculin skin testing have limitations and are thus not always helpful in diagnosing TB. Smear microscopy alone, although cheap and easy to perform, has both the problems of sensitivity and specificity [2]. Although the culture is more sensitive than the smear microscopy, the time required, automated liquid culture systems like BACTEC or Mycobacteria Growth Indicator Tube (MGIT) take 1–6 weeks for growth detection, and frequent negative results in paucibacillary specimens are important limitation [2], [3], [4] and need specialized laboratory personnel [5]. This leads to a diagnostic delay that impedes disease control, and increases healthcare costs [1]. In the last few years, since the discovery of the polymerase chain reaction(PCR), a large number of molecular techniques have been developed [6]. However, their sensitivity is greatly dependent on the efficiency of the sample preparation, DNA extraction and the presence of PCR inhibitors [7], [8] and such methods require expertise and highly sophisticated settings that are not cost-effective for the laboratory in low-resource countries. Therefore, a simple, rapid and effective method for TB diagnosis remains to be developed.
One of latest assay, loop-mediated isothermal amplification(LAMP, Eiken Chemical Co Ltd., Tokyo, Japan) assay has been developed as a novel technique for nucleic acid amplification [9] and was evaluated in large studies recently [10]. LAMP can amplify DNA with high specificity and efficiency under isothermal conditions. Unlike PCR, LAMP reaction does not require a denaturated DNA template and relies on auto cycling strand displacement DNA synthesis by a Bst DNA polymerase [11]. The large amount of DNA generated in less than an hour and positive LAMP reaction can be visualized with the naked eye, without need for gel electrophoresis [12].
The diagnostic performance of LAMP assay has been investigated in several studies, which have variable results. The aim of this meta-analysis is to establish the overall diagnostic accuracy of LAMP assay and identify potential confounders or effective modifiers of its value in pulmonary tuberculosis (TB), thus providing the important up-to-date information on LAMP assay for pulmonary TB diagnosis.
Section snippets
Search strategy
We identified studies that showed the evidence of using LAMP assay in order to diagnose pulmonary TB. Two investigators (Y.L. and L.Y.) independently performed a systematic electronic search of the pubmed, embase databases and the cochrane-controlled central register of controlled trials and included reports for original articles published until 1 October 2012 to identify potentially relevant articles and abstracts. The following search terms were used: “LAMP Assay” OR “LAMP” OR “loop-mediated
Literature search and study selection
A total of 135 studies were screened for analysis of patients with tuberculosis using the novel and sensitive LAMP assay. After full-text review, 10 studies eventually met the inclusion criteria (Fig. 1).
Characters of the selected studies
Overall, the selected 10 studies [18], [19], [20], [21], [22], [23], [24], [25], [26], [27] included 1920 suspected specimens of having pulmonary TB, which included 823 pulmonary TB specimens (Patients with microbiologically confirmed TB), and 1097 non-TB specimens (The samples of patients were
Discussion
In this study, we clarified the diagnostic accuracy of LAMP assay for pulmonary TB. In addition, the diagnostic accuracy in different quality levels of studies were also explored. To our knowledge, this is the first systematic review and meta-analysis to estimate the diagnostic accuracy of LAMP assay in pulmonary TB.
The results of these 10 studies showed that LAMP assay was accurate to diagnose pulmonary TB. Using the bivariate random-effects approach, we found a summary AUC of 0.9633, a
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgments
Conceived and designed the experiments:YL, LY. Performed the experiments:MW, WX, ZK. Analyzed the data:YL.Wrote the paper: YL, LY. This work was supported by a grant from the National Clinical Key Specialty Construction Projects to the Department of Clinical Laboratory of Renmin Hospital of Wuhan University.
References (32)
- et al.
Modern approaches to a rapid diagnosis of tuberculosis: promises and challenges ahead
Tuberculosis (Edinb)
(2007) - et al.
Isolation of single-stranded DNA from loop-mediated isothermal amplification products
Biochem Biophys Res Commun
(2002) - et al.
Application of loop-mediated isothermal amplification technique to rapid and direct detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood cultures
J Infect Chemother
(2007) - et al.
Accelerated reaction by loop-mediated isothermal amplification using loop primers
Mol Cell Probes
(2002) - et al.
Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation
Biochem Biophys Res Commun
(2001) - et al.
Evaluation of reverse transcription loop-mediated isothermal amplification in conjunction with ELISA-hybridization assay for molecular detection of Mycobacterium tuberculosis
J Microbiol Methods
(2009) - et al.
Use of visual loop-mediated isotheral amplification of rimM sequence for rapid detection of Mycobacterium tuberculosis and Mycobacterium bovis
J Microbiol Methods
(2009) - et al.
Diagnostic accuracy of nucleic acid amplification tests for tuberculous meningitis: a systematic review and meta-analysis
Lancet Infect Dis
(2003) Global tuberculosis control: WHO report
(2012)- et al.
Simplified detection of Mycobacterium tuberculosis in sputum using smear microscopy and PCR with molecular beacons
J Med Microbiol
(2007)
Newer diagnostic techniques for tuberculosis
Indian J Med Res
Mycobacterial growth and bacterial contamination in the mycobacteria growth indicator tube and BACTEC 460 culture systems
J Clin Microbiol
Molecular diagnostics in tuberculosis
Eur J Clin Microbiol Infect Dis
Rapid diagnosis of extrapulmonary tuberculosis by PCR: impact of sample preparation and DNA extraction
J Clin Microbiol
Diagnosis of extrapulmonary tuberculosis by smear, culture, and PCR using universal sample processing technology
J Clin Microbiol
Development and validation of methods for assessing the quality of diagnostic accuracy studies
Health Technol Assess
Cited by (22)
Loop-mediated isothermal amplification (LAMP) in the respiratory specimens for the diagnosis of pediatric pulmonary tuberculosis: A pilot study
2020, Journal of Infection and ChemotherapyCitation Excerpt :For similar reasons it scores over other tests for paucibacillary TB, especially in pediatric population. Various studies on adults with TB have demonstrated LAMP to have higher sensitivity and specificity [16–30]. There is limited literature on the usage of LAMP in the diagnosis of childhood TB [31].
Diagnostic scoring system for tuberculous meningitis among adult patients with non-suppurative and non-bacterial meningitis
2018, Journal of Infection and ChemotherapyCitation Excerpt :However, these techniques are unpractical due to the relatively high cost and the requirement for experienced technicians. In addition, these techniques have limited data on the diagnosis of TBM [10–12]. Adenosine deaminase (ADA) is an enzyme involved in purine metabolism.
Systematic review: Comparison of Xpert MTB/RIF, LAMP and SAT methods for the diagnosis of pulmonary tuberculosis
2016, TuberculosisCitation Excerpt :The I2 values for sensitivity and specificity were >80% for all three of the diagnostic tests, suggesting that significant heterogeneity existed between the studies of each, which prevented us from pooling the data. Meta-analysis of LAMP for the diagnosis of PTB [16], found that the I2 values for sensitivity and specificity were 97.1% and 86.7%, respectively, but significant heterogeneity clearly existed after subgroup analysis. In a previous meta-analysis of Xpert MTB/RIF [20], the I2 values for the sensitivity and specificity for the diagnosis of smear-negative PTB were 70.8% and 69.7%, respectively.
TB-ISATEST: a Diagnostic LAMP Assay for Differentiation of Mycobacterium tuberculosis
2023, Russian Journal of Bioorganic ChemistryCombining LAMP and Au-Nanoprobe to detect INH-RIF resistance accurately in tuberculosis: An evidence-based review
2021, Journal of Infection in Developing Countries
- 1
These authors contributed equally to this work.