Development of asymmetric hairpins-mediated nucleic acid isothermal amplification-based lateral flow detection of Mycobacterium tuberculosis

2022 ◽  
Vol 350 ◽  
pp. 130836
Author(s):  
Hongbo Shan ◽  
Yang Wang ◽  
Tao Wu ◽  
Binwu Ying ◽  
Gaolian Xu
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Flow Detection

scholarly journals Clinical profiling and use of loop-mediated isothermal amplification assay for rapid detection of Mycobacterium tuberculosis from sputum

1970 ◽  
Vol 7 (2) ◽  
pp. 109-114 ◽  
Author(s):  
A Poudel ◽  
BD Pandey ◽  
B Lekhak ◽  
B Rijal ◽  
BR Sapkota ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Common Symptom ◽  
Bcg Vaccination ◽  
Loop Mediated Isothermal Amplification ◽  
16S Rna ◽  
Clinical Profiling

Background: Tuberculosis is a global health problem and the situation is worsening with newer incidences of drug resistance and HIV association. Diagnosis of tuberculosis can be done by many methods and test, culture of sputum being the ideal one. Nucleic acid amplification (NAA) assay are more time efficient one, that amplify and detect specific nucleic acid sequences allows rapid, sensitive and specific detection of M. tuberculosis in sputum samples. Objectives: The present study intends to compile the clinical presentations of the pulmonary tuberculosis (PTB) patients and to evaluate the efficacy of in-house loop-mediated isothermal amplification (LAMP) in detecting Mycobacterium tuberculosis in sputum samples by comparing with microscopy and culture. Materials and methods: Two hundred two sputum samples were collected from 202 patients at National Tuberculosis Center, Bhaktapur, Nepal. Complete clinical profiling, epidemiological data and record on BCG vaccination were noted and the samples were subjected for microscopy, culture and in-house LAMP with six primers specific for 16S RNA gene of Mycobacterium tuberculosis. Result: Of the 176 cases of clinical profiling, productive cough was most common symptom in 147 (83.52%), followed by chest pain 136 (77.27%), fever 133 (75.56%) and haemoptysis 61 (34.66%). There was a statistically significant association between BCG vaccination and development of TB (Χ2=5.33, P=0.02). Of 202 cases, 115 (56.93%) were chest X-ray positive, 101(50%) were direct smear-positive and 100 (49.51%) were culture positive. LAMP had a sensitivity of 97% and specificity of 94.12% while comparing with culture. In addition, its sensitivity and specificity were 91.09% and 89.11% respectively with reference to microscopy. Conclusion: As in our previous study, overall, the result of present study further confirms that the in-house LAMP is a simple, rapid, sensitive and specific DNA amplification technique for PTB diagnosis. Because of rapidity of microscopy and specificity of culture, in-house LAMP assay can be used as a very powerful and useful supplementary tool with complete clinical profiling of the patients for rapid diagnosis of TB in both AFB-positive and negative cases who are suspected as PTB in disease endemic country like Nepal. Key words: clinical profiling; Sputum; DNA; LAMP; M. tuberculosis; Nepal DOI: 10.3126/kumj.v7i2.2701 Kathmandu University Medical Journal (2009) Vol.7, No.2 Issue 26, 109-114

scholarly journals Rapid and Visual Differentiation of Mycobacterium tuberculosis From the Mycobacterium tuberculosis Complex Using Multiplex Loop-Mediated Isothermal Amplification Coupled With a Nanoparticle-Based Lateral Flow Biosensor

2021 ◽  
Vol 12 ◽  
Author(s):  
Xinggui Yang ◽  
Junfei Huang ◽  
Xu Chen ◽  
Ziyu Xiao ◽  
Xiaojuan Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Limit Of Detection ◽  
Clinical Specimen ◽  
Lamp Assay ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Whole Process ◽  
Specimen Processing ◽  
Mycobacterium Africanum

Tuberculosis (TB) is a chronic infectious disease mainly caused by Mycobacterium tuberculosis (MTB), but other members of the Mycobacterium tuberculosis complex (MTBC), especially Mycobacterium bovis (pyrazinamide-resistant organisms), may also be involved. Thus, the ability to rapidly detect and identify MTB from other MTBC members (e.g., M. bovis, Mycobacterium microti, Mycobacterium africanum) is essential for the prevention and treatment of TB. A novel diagnostic method for the rapid detection and differentiation of MTB, which employs multiplex loop-mediated isothermal amplification (mLAMP) combined with a nanoparticle-based lateral flow biosensor (LFB), was established (mLAMP-LFB). Two sets of specific primers that target the IS6110 and mtp40 genes were designed according to the principle of LAMP. Various pathogens were used to optimize and evaluate the mLAMP-LFB assay. The optimal conditions for mLAMP-LFB were determined to be 66°C and 40 min, and the amplicons were directly verified by observing the test lines on the biosensor. The LAMP assay limit of detection (LoD) was 125 fg per vessel for the pure genomic DNA of MTB and 4.8 × 103 CFU/ml for the sputum samples, and the analytical specificity was 100%. In addition, the whole process, including the clinical specimen processing (35 min), isothermal amplification (40 min), and result confirmation (1–2 min), could be completed in approximately 80 min. Thus, mLAMP-LFB is a rapid, reliable, and sensitive method that is able to detect representative members of MTBC and simultaneously differentiate MTB from other MTBC members, and it can be used as a potential screening tool for TB in clinical, field, and basic laboratory settings.

scholarly journals Application and research of loop-mediated isothermal amplification and lateral flow dipstick in detection of Porcine circovirus type 3

2019 ◽  
Author(s):  
Yan Wang ◽  
Xianjie Han ◽  
Jianli Shi ◽  
Zhe Peng ◽  
Xiaoyan Wu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Acid Concentration ◽  
Clinical Symptoms ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Lateral Flow Dipstick ◽  
Nucleic Acid Concentration ◽  
Pcr Method ◽  
Type 3

Abstract Background：Porcine circovirus type 3 (PCV3) is a newly detected pathogen from pigs in the past few years. As the infection rate of PCV3 in the herd is getting higher and wider, the infection is more and more serious. The virus with unclear clinical symptoms and pathogenesis is also paid more and more attention. And detection of pathogenic antibodies has become an important issue. Methods:In this experiment, Loop-mediated isothermal amplification (LAMP) was combined with Lateral flow dipstick (LFD) to establish a rapid and convenient detection method (LAMP-LFD). Results:The results showed that the band was darkened at a nucleic acid concentration of 0.2 pg/ μL using a conventional PCR method, and a clear detection line was observed at a nucleic acid concentration of 0.2 fg/μL using the LAMP-LFD method. The primer and hybridization probe are only compatible with PCV3.Compared to the PCR method, the LAMP-LFD method has a shorter time (about 1 h) and requires less instrumentation. Conclusions: The test results show that the LAMP-LFD method has extremely high sensitivity and specificity. The operation is simple and quick. The visual effect of the detection results is better than that of PCR and single LAMP test, which can bring convenience in practical production applications.

Proficient Detection of Multi-Drug-Resistant Mycobacterium tuberculosis by Padlock Probes and Lateral Flow Nucleic Acid Biosensors

2016 ◽  
Vol 88 (8) ◽  
pp. 4277-4284 ◽  
Author(s):  
Asalapuram R Pavankumar ◽  
Anna Engström ◽  
Jie Liu ◽  
David Herthnek ◽  
Mats Nilsson
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Lateral Flow ◽  
Drug Resistant ◽  
Padlock Probes

Development and application of loop-mediated isothermal amplification label-based nanoparticles lateral flow biosensor for detection of Mycobacterium tuberculosis

2019 ◽  
Author(s):  
Xingyun Wang ◽  
Yi Wang ◽  
Weiwei Jiao ◽  
Guirong Wang ◽  
Yacui Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Diagnostic Technique ◽  
Cost Effective ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
High Morbidity ◽  
Analytical Sensitivity ◽  
Loop Mediated Isothermal Amplification

Abstract Tuberculosis is a serious disease with high morbidity and mortality, thus rapid and cost-effective diagnostic test for Mycobacterium tuberculosis (MTB) is urgently needed. Here, a novel detection diagnostic technique, termed as loop-mediated isothermal amplification label-based nanoparticles with lateral flow biosensor (LAMP-LFB), was developed and evaluated for rapid, reliable and objective detection of MTB. Two sets of primers, which targeted IS 6110 and IS 1081 sequences of MTB, were simultaneously designed for establishment of LAMP-LFB assay. The optimal reaction conditions of MTB-LAMP-LFB assay confirmed were 66ºC for only 50min. The analytical sensitivity of MTB-LAMP-LFB is 10fg of genomic templates in pure culture, and the detection results obtained from LFB was in conformity with agarose gel electrophoresis. No cross-reactivity with other common bacteria and non-tuberculous mycobacteria strains (NTM) was obtained. A total of 158 clinical samples were collected from presumptive 158 TB patients, were used for evaluating the feasibility of MTB-LAMP-LFB assay. Among 98 TB patients diagnosed with composite reference standard, the positive rate for MTB detection using liquid culture, Xpert MTB/RIF and LAMP-LFB were 40.0% (39/98), 50.0% (48/98), and 86.7% (85/98), respectively. Among 39 culture confirmed samples, 84.6% (33/39) cases were Xpert MTB/RIF-positive and 92.3% (36/39) were LAMP-LFB-positive. For the 59 clinically diagnosed TB cases 25.4% (15/59) and 83.0% (49/59) were Xpert MTB/RIF-positive and LAMP-LFB positive, respectively. Therefore, MTB-LAMP-LFB assay is a simple, reliable, and sensitive method for MTB detection and maybe prospective in early diagnosis of MTB.

On-point detection of GM rice in 20 minutes with pullulan as CPA acceleration additive

2014 ◽  
Vol 6 (23) ◽  
pp. 9198-9201 ◽  
Author(s):  
Rui Wang ◽  
Jian Wu ◽  
Fang Zhang ◽  
Liu Wang ◽  
Feng Ji
Keyword(s):  
Nucleic Acid ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Specific Detection ◽  
Lateral Flow Dipstick ◽  
Gm Rice ◽  
Point Detection

1% (w/v) concentration of pullulan as a cross-priming isothermal amplification additive has an acceleration effect on nucleic acid amplification. With this property, on-point specific detection of DNA by lateral-flow dipstick within 20 minutes was realized.

scholarly journals Integrated nucleic acid testing system to enable TB diagnosis in peripheral settings

Lab on a Chip ◽  
2020 ◽  
Vol 20 (21) ◽  
pp. 4071-4081
Author(s):  
Hsiang-Wei Lu ◽  
Rama Sakamuri ◽  
Pranav Kumar ◽  
Tanya M. Ferguson ◽  
Robert W. Doebler ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Dna Extraction ◽  
Dna Amplification ◽  
Lateral Flow ◽  
Testing System ◽  
Nucleic Acid Testing ◽  
Testing Device ◽  
Flow Detection

We developed a nucleic acid testing device that automates pathogen lysis, DNA extraction, isothermal DNA amplification and lateral flow detection.

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