Utility of loop-mediated isothermal amplification as a point-of-care test in diagnosis of amoebic liver abscess

2021 ◽  
pp. 004947552110180
Author(s):  
Dipti Handa ◽  
Monica Gupta ◽  
Sarabmeet Singh Lehl ◽  
Amit Gupta ◽  
Ram Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Liver Abscess ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Amoebic Liver Abscess ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Loop Mediated Isothermal Amplification ◽  
Polymerase Chain

Definitive diagnosis of amoebic liver abscess is challenging owing to the unavailability of sensitive commercial point-of-care molecular tests. The primary aim of our prospective diagnostic study was to compare available laboratory methods for the diagnosis of Entamoeba histolytica in clinical samples with loop-mediated isothermal amplification. We compared deoxyribonucleic acid (DNA) analysis methods, namely, loop-mediated isothermal amplification and reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) using pus, stool and blood samples from 200 patients with clinical and radiological diagnosis of amoebic liver abscess. Loop-mediated isothermal amplification had significantly higher sensitivity (88%) as compared to reverse transcriptase polymerase chain reaction (64%) and excellent specificity (100%).

scholarly journals Loop-Mediated Isothermal Amplification (LAMP) as a Promising Point-of-Care Diagnostic Strategy in Avian Virus Research

Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 76
Author(s):  
Faiz Padzil ◽  
Abdul Razak Mariatulqabtiah ◽  
Wen Siang Tan ◽  
Kok Lian Ho ◽  
Nurulfiza Mat Isa ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Point Of Care ◽  
Rolling Circle Amplification ◽  
Isothermal Amplification ◽  
Screening Methods ◽  
Diagnostic Strategy ◽  
Chain Reaction ◽  
Loop Mediated Isothermal Amplification ◽  
Polymerase Chain

Over the years, development of molecular diagnostics has evolved significantly in the detection of pathogens within humans and their surroundings. Researchers have discovered new species and strains of viruses, while mitigating the viral infections that occur, owing to the accessibility of nucleic acid screening methods such as polymerase chain reaction (PCR), qualitative (real-time) polymerase chain reaction (qPCR) and reverse-transcription qPCR (RT-qPCR). While such molecular detection methods are widely utilized as the benchmark, the invention of isothermal amplifications has also emerged as a reliable tool to improvise on-field diagnosis without dependence on thermocyclers. Among the established isothermal amplification technologies are loop-mediated isothermal amplification (LAMP), recombinant polymerase amplification (RPA), strand displacement activity (SDA), nucleic acid sequence-based amplification (NASBA), helicase-dependent amplification (HDA) and rolling circle amplification (RCA). This review highlights the past research on and future prospects of LAMP, its principles and applications as a promising point-of-care diagnostic method against avian viruses.

Comparison of Reverse Transcriptase Loop-Mediated Isothermal Amplification and Reverse Transcriptase Polymerase Chain Reaction for Detection of Prostate Specific Antigen

2019 ◽  
Author(s):  
Mohammad Amin Almasi ◽  
Marya Esmaili
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Isothermal Amplification ◽  
Chain Reaction ◽  
Loop Mediated Isothermal Amplification ◽  
Polymerase Chain ◽  
Positive Controls

Background: Research shows that prostate cancer ranks second among the top five most common cancers in men. It has been confirmed that when circulating Prostate Specific Antigen (PSA) transcripts are successfully detected, prostate cancer cells can be diagnosed at an early stage. A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed and compared to reverse transcriptase polymerase chain reaction (RT-PCR) assay for detection of PSA. Methods: 47 patients, including 30 patients with prostate cancer, 15 with Benign Prostate Hyperplasia (BPH) and 2 healthy subjects as negative controls were included in this study. The prostate cancer cell lines (PC3 and LNCaP) of two patients were included in the study as positive controls. Next, RNA was extracted from fresh samples and a first strand cDNA synthesis kit was applied for the synthesis of cDNA. The human prostate specific antigen gene was used to design specific primers. Results: The results indicated that the control subjects and participants suffering from BPH were not positive. 13 out of 15 (86.6%) patients suffering from localized cancer were PSA positive. PSA positive results were observed among all 15 metastatic patients and positive controls (100%). RT-LAMP is an advantageous method because it is highly sensitive (1000-fold), quite cheap, user-friendly, and safe; in addition, it can be quickly performed by visual detection using GineFinderTM dye in a water bath. Conclusion: RT-LAMP technique can be simply and reliably applied with the aid of basic instruments, and its results can be visually inspected in laboratory studies.

scholarly journals Detecting SARS-CoV-2 at point of care: preliminary data comparing loop-mediated isothermal amplification (LAMP) to polymerase chain reaction (PCR)

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Marc F. Österdahl ◽  
Karla A. Lee ◽  
Mary Ni Lochlainn ◽  
Stuart Wilson ◽  
Sam Douthwaite ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Predictive Value ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Service Improvement ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Loop Mediated Isothermal Amplification ◽  
Polymerase Chain

Abstract Background A cost effective and efficient diagnostic tool for COVID-19 as near to the point of care (PoC) as possible would be a game changer in the current pandemic. We tested reverse transcription loop mediated isothermal amplification (RT-LAMP), a method which can produce results in under 30 min, alongside standard methods in a real-life clinical setting. Methods This prospective service improvement project piloted an RT-LAMP method on nasal and pharyngeal swabs on 21 residents of a high dependency care home, with two index COVID-19 cases, and compared it to multiplex tandem reverse transcription polymerase chain reaction (RT-PCR). We recorded vital signs of patients to correlate clinical and laboratory information and calculated the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of a single swab using RT-LAMP compared with the current standard, RT-PCR, as per Standards for Reporting Diagnostic Accuracy Studies (STARD) guidelines. Results The novel method accurately detected 8/10 RT-PCR positive cases and identified a further 3 positive cases. Eight further cases were negative using both methods. Using repeated RT-PCR as a “gold standard”, the sensitivity and specificity of a single novel test were 80 and 73% respectively. PPV was 73% and NPV was 83%. Incorporating retesting of low signal RT-LAMP positives improved the specificity to 100%. We also speculate that hypothermia may be a significant early clinical sign of COVID-19. Conclusions RT-LAMP testing for SARS-CoV-2 was found to be promising, fast and to work equivalently to RT-PCR methods. RT-LAMP has the potential to transform COVID-19 detection, bringing rapid and accurate testing to the PoC. RT-LAMP could be deployed in mobile community testing units, care homes and hospitals to detect disease early and prevent spread.

scholarly journals LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR DETECTION OF C. BURNETII IN BULGARIA

2021 ◽  
Vol 19 (2) ◽  
pp. 147-151
Author(s):  
M. Kunchev ◽  
V. Belcheva ◽  
E. Grigorov
Keyword(s):  
Polymerase Chain Reaction ◽  
Sensitivity And Specificity ◽  
Q Fever ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Chain Reaction ◽  
Retrospective Diagnosis ◽  
Loop Mediated Isothermal Amplification ◽  
The World ◽  
Polymerase Chain

Q fever, which is caused by Coxiella burnetii, a small, pleomorphic intracellular bacterium, is the most widespread zoonosis in the world. The chronic form of the disease can lead to disability and death. Rapid diagnosis of Q fever is needed in order that effective treatment can be initiated. The conventional retrospective diagnosis of Q fever, based on serology, is useless for the treatment of afflicted patients. Thus, molecular methods have been created to close the diagnostic gap between the onset of the disease and the presence of specific antibodies in serum. A polymerase chain reaction is a suitable and reliable method with high sensitivity and specificity, but it requires expensive equipment and post-amplification protocol. Loop-mediated isothermal amplification (LAMP) is an isothermal technique, conducted at constant temperature that can amplify a negligible amount of DNA to more than 109 copies within one hour, using special primers and polymerase. We have tested the sensitivity and specificity of LAMP in the detection of C. burnetii. The mean positive rate of LAMP and polymerase chain reaction in patients was 100% and 74%, respectively. LAMP reacted negatively with non-C. burnetii pathogens and non-infected blood samples. We conclude that LAMP is a sensitive and specific technique for the detection of C. burnetii and has advantages over serological methods and PCR that make it attractive for diagnosing Q fever in countries around the world.

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