Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis

Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui; Hu, Guohua

doi:10.1371/journal.pone.0186985

Cited by 17 publications

(15 citation statements)

References 26 publications

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“…As there are many limitations in microbiological tests, new approaches were developed for detection of M. Tb [49][50][51][52]. Compared with Ziehl-Neelsen staining and culture, IGRAs break new ground in TBM diagnosis as they are much more rapid and simple.…”

Section: Risk Of Bias Applicability Concernsmentioning

confidence: 99%

Interferon-γ release assays for tuberculous meningitis diagnosis: a meta-analysis

et al. 2021

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IntroductionTuberculous meningitis (TBM) is still a great challenge to global public health. As conventional diagnostic methods for TBM are unsatisfactory, interferon-γ release assays (IGRAs) have been introduced for TBM diagnosis tentatively. However, the role of IGRAs for diagnosing TBM remains unclear. Thus, we systematically evaluated the diagnostic performance of cerebrospinal fluid (CSF) and peripheral blood (PB) IGRAs in TBM to fill this blank.Material and methodsRelevant studies were systematically searched in both foreign and Chinese databases up to March 2018. Studies in which TBM diagnosis was based on microbiological or clinical criteria were included. The quality of the included studies was assessed through the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) tool. Main outcome measures, including sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR) and diagnostic odds ratio (DOR), were pooled statistically using random effects models. The potential heterogeneity was explored by threshold effect analysis, subgroup analyses and meta-regression. Funnel plots and Egger’s test were used to test the potential publication bias. Statistical analyses were performed using Stata and Meta-DiSc software.ResultsTwenty-six out of 656 publications were eligible for meta-analysis, including 1892 participants in total. The pooled estimates of PB IGRAs for TBM diagnosis are as follows: sensitivity: 0.81 (95% CI: 0.78–0.84); specificity: 0.76 (95% CI: 0.73–0.78); PLR: 4.23 (95% CI: 2.95–6.07); NLR: 0.24 (95% CI: 0.19–0.32) and DOR: 21.06 (11.91-37.24). The corresponding estimates for CSF IGRAs were obtained: sensitivity: 0.81 (95% CI: 0.76–0.85); specificity: 0.89 (95% CI: 0.86–0.92); PLR: 7.87 (95% CI: 4.98–12.46); NLR: 0.19 (95% CI: 0.13–0.29); and DOR: 47.74 (25.02–91.12).ConclusionsThe diagnostic performance of IGRAs is suboptimal. In terms of cost, turn-around time and accessibility, these assays are unsuitable for use as biomarkers for TBM diagnosis.

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Section: Risk Of Bias Applicability Concernsmentioning

confidence: 99%

Interferon-γ release assays for tuberculous meningitis diagnosis: a meta-analysis

et al. 2021

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“…PCR-LFA is a method that combines PCR and LFA to detect PCR products rapidly. The PCR products are double-labeled by gene-specific primer sets to generate sandwich assays [19,20]. Detection is performed by trapping the labeled PCR products on the surface of a lateral flow strip possessing test and control lines [20,22].…”

Section: Introductionmentioning

confidence: 99%

“…This methodology is simpler than conventional PCR-gel electrophoresis, requiring minimal personnel training. Also, PCR-LFA allows for point-of-care testing, allowing faster and easier detection of pathogens in the field environment [19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Rapid diagnosis of two marine viruses, red sea bream iridovirus and viral hemorrhagic septicemia virus by PCR combined with lateral flow assay

et al. 2020

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Red sea bream iridovirus (RSIV) and Viral hemorrhagic septicemia virus (VHSV) are the most important viral marine pathogens in South Korea because RSIV and VHSV infect and cause high mortality rates in major fish species such as Paralichthys olivaceus and Sebastes schlegelii. These viruses can be transmitted both vertically and horizontally, and early diagnosis is imperative. In this research, RSIV and VHSV viral genomes are detected by PCR-lateral flow assay (LFA). PCR-LFA is sensitive, capable of detecting a viral genome at a concentration of 2-200 fg/lL. Development of this detection method is very meaningful because LFA is simple, requiring a minimum of personnel training to perform. Additionally, LFA requires less time than other detection methods and can be an immediate detection tool that is indispensable in preventing rapid viral spread.

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“…Ma et al combined a LFTS with recombinase polymerase amplification for detection of MTB complex [32]. Using the IS6110 sequence of tuberculous meningitis as a target, fragments were amplified by nested PCR and two sets of primers, and then detected by an antibody-labeled probe on a LFTS [33]. A PCR-nucleic acid lateral flow immunoassay was also reported for the indirect de-tection of labeled PCR amplification products of multidrug-resistant TB [34].…”

Section: Introductionmentioning

confidence: 99%

Lateral Flow Genochromatographic Strip for Naked-Eye Detection of Mycobacterium Tuberculosis PCR Products with Gold Nanoparticles as a Reporter

Nazari-Vanani

Tondro

et al. 2020

jbpe

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Background: Mycobacterium tuberculosis (MTB) is a pathogen causing tuberculosis (TB) in human, and TB can cause enormous social and economic disruptions. Lateral flow test strips (LFTSs) are inexpensive, portable, disposable, rapid, and easy-to-use analytical tools.Objective: LFTSs were prepared for the detection of MTB. LFTSs were fabricated using a new specific probe for MTB H37Rv, based on IS6110 sequence gene, and tailed with poly deoxyadenine (dA). Material and Methods:In this experimental study, to create test and control zones, streptavidin (STP) and a 150-mer dA were dotted on a nitrocellolose membrane. Gold nanoparticles (GNPs) were conjugated with poly deoxythymidine sequence and placed on the conjugate pad. The composition of immersion buffers for sample pad and conjugate pad, running solution, solutions of GNPs-S-dT150 and STP were introduced. DNA genome of MTB and Mycobacterium bovis in clinical samples was amplified with PCR, and then detected by the LFTSs. During the assay, samples were firstly hybridized in two steps and then placed on a conjugate pad in a manner that positive and negative samples provided two and one red lines, respectively, on the detection pad.Results: After PCR reaction with biotinylated primer, hybridization process with specific MTB probe-dA70-100 toke 10 min, and running process on the strip was performed within 5 min. Conclusion:We showed that LFTS can discriminate a particular bacteria strain from others. The LFTSs can be redesigned for detection of other pathogenic genomes.

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Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis

Cited by 17 publications

References 26 publications

Interferon-γ release assays for tuberculous meningitis diagnosis: a meta-analysis

Interferon-γ release assays for tuberculous meningitis diagnosis: a meta-analysis

Rapid diagnosis of two marine viruses, red sea bream iridovirus and viral hemorrhagic septicemia virus by PCR combined with lateral flow assay

Lateral Flow Genochromatographic Strip for Naked-Eye Detection of Mycobacterium Tuberculosis PCR Products with Gold Nanoparticles as a Reporter

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