scholarly journals Diagnostic performance of a novel loop-mediated isothermal amplification (LAMP) assay targeting the apicoplast genome for malaria diagnosis in a field setting in sub-Saharan Africa

2015 ◽  
Vol 14 (1) ◽  
Author(s):  
Eniyou C. Oriero ◽  
Joseph Okebe ◽  
Jan Jacobs ◽  
Jean-Pierre Van geertruyden ◽  
Davis Nwakanma ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Lamp Assay ◽  
Malaria Diagnosis ◽  
Isothermal Amplification ◽  
Sub Saharan Africa ◽  
Loop Mediated Isothermal Amplification ◽  
Sub Saharan ◽  
Field Setting ◽  
Apicoplast Genome

scholarly journals Evaluation of Loop Mediated Isothermal Amplification (LAMP) for Diagnosis of Plasmodium falciparum in Wad Medani City, Gezira State, Sudan

2019 ◽  
pp. 1-7
Author(s):  
M. Y. Mohamed ◽  
A. D. Abakar ◽  
B. A. Talha ◽  
Salah Eldin G. Elzaki ◽  
Y. A. Mohammed ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Falciparum Malaria ◽  
Diagnostic Performance ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Teaching Hospitals ◽  
Molecular Technique ◽  
Rrna Gene ◽  
Predictive Values ◽  
Loop Mediated Isothermal Amplification

Plasmodium falciparum considered as the most serious form of species causes malaria compared with other species. Diagnosis of falciparum malaria in Sudan remain a major problem, the laboratory diagnosis depends solely on microscopy and RDTs. Loop mediated isothermal amplification (LAMP) assay is a molecular technique done in isothermal temperature using simple, inexpensive instruments for detection of falciparum malaria. The aim of the study is to evaluate the diagnostic performance of loop mediated isothermal amplification (LAMP) assay for detection of P. falciparum and compare with microscopic detection. A cross sectional hospital based study conducted on 220 blood samples collected from participants suspected to have falciparum malaria attending Wad Medani Teaching Hospitals and 26 healthy participants during the period November 2018 to January 2019. Thick blood films were done and used for P. falciparum detection. The extracted DNA by TE buffer was amplified by LAMP assay targeting 18S rRNA gene. Data were analyzed using Medical calculator (MedCalc) programs (V. 16). The results showed that the sensitivity, specificity, positive predictive value, negative predictive values were 99.1%, 84.6%, 53.2%, 99.8% respectively. Validation of LAMP diagnostic performance revealed that area under the curve is 0.919, while Weighted Kappa is 0.866. The study concluded that the LAMP assay had the identical diagnostic performance compared with microscopy in diagnosis of Plasmodium falciparum malaria. This gives a relative effortlessness application of LAMP assay in Sudan after availing the required logistics.

scholarly journals Comparative Diagnostic Performance of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Kit for the Rapid Detection of SARS-CoV-2

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1629
Author(s):  
Alexander Domnich ◽  
Andrea Orsi ◽  
Donatella Panatto ◽  
Vanessa De Pace ◽  
Valentina Ricucci ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Diagnostic Performance ◽  
Point Of Care ◽  
Lamp Assay ◽  
Laboratory Diagnosis ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Rt Pcr ◽  
Laboratory Equipment ◽  
Loop Mediated Isothermal Amplification

Although the reverse transcription-polymerase chain reaction (RT-PCR) is considered a standard-of-care assay for the laboratory diagnosis of SARS-CoV-2, several limitations of this method have been described. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an alternative molecular assay and is potentially able to overcome some intrinsic shortcomings of RT-PCR. In this study, we evaluated the diagnostic performance of the novel HG COVID-19 RT-LAMP assay. In this retrospective analysis, a total of 400 routinely collected leftover nasopharyngeal samples with a known RT-PCR result were tested by means of the HG COVID-19 RT-LAMP assay. The overall sensitivity and specificity values of HG COVID-19 RT-LAMP versus RT-PCR were 97.0% (95% CI: 93.6–98.9%) and 98.5% (95% CI: 95.7–99.7%), respectively. Inter-assay agreement was almost perfect (κ = 0.96). Concordance was perfect in samples with high viral loads (cycle threshold < 30). The average time to a positive result on RT-LAMP was 17 min. HG COVID-19 RT-LAMP is a reliable molecular diagnostic kit for detecting SARS-CoV-2, and its performance is comparable to that of RT-PCR. Shorter turnaround times and the possibility of performing molecular diagnostics in the point-of-care setting make it a valuable option for facilities without sophisticated laboratory equipment.

scholarly journals Diagnostic Performance of Blood Film Microscopy and PfHRP2-based RDT in a Routine Clinical Setting of a Secondary Health Facility in Ghana

2019 ◽  
pp. 1-9
Author(s):  
Francis O. Agyapong ◽  
Daniel Ansong ◽  
Alex Owusu-Ofori ◽  
Ruby Martin-Peprah
Keyword(s):  
Diagnostic Performance ◽  
Malaria Diagnosis ◽  
Blood Film ◽  
Molecular Techniques ◽  
Sub Saharan Africa ◽  
Reference Test ◽  
Paediatric Age ◽  
Sub Saharan ◽  
Low Sensitivity ◽  
High Level

Background: Malaria remains a major public health threat claiming many lives particularly in Sub-Saharan Africa. Light microscopy and RDT are the mainstay tests in the clinical settings for malaria diagnosis. Many studies report varying levels of validity of these tests compared to molecular methods like PCR. Documentation on such comparative study involving the use of molecular techniques as reference test is scanty in Ghana. This study therefore assesses the diagnostic performance of these tests compared to PCR. Methods: Blood film microscopy (thin and thick), RDT and nested PCR were run on blood samples from a total of 188 malaria suspected patients. The accuracy indices of the microscopy and RDT were calculated using the results of the PCR as the reference test. Results: A total of 188 patients were recruited with females constituting the majority 128 (68%). The paediatric age group 1-10 years carried the largest burden of malaria by means of all the 3 tests. A sensitivity of 47.37% (95% ci, 37.03 – 57.88%) was shown by both the microscopy and RDT with specificity of 93.55% (95% ci, 86.48 – 97.60) and 100% (95% ci, 96.11 – 100.00%) and kappa co – efficient of 0.41 and 0.47 respectively. Conclusion: Both microscopy and RDT exhibited high level of specificity but low sensitivity. Significant number of malaria parasitaemic patients as revealed by the PCR was missed by both the RDT and blood film microscopy and thus went undiagnosed.

Development and evaluation of a loop-mediated isothermal amplification (LAMP) diagnostic test for detection of whipworm, Trichuris trichiura, in faecal samples

2020 ◽  
Vol 94 ◽  
Author(s):  
M.G. Ngari ◽  
I.N. Mwangi ◽  
M.P. Njoroge ◽  
J. Kinyua ◽  
F.A. Osuna ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Infection Intensity ◽  
Lamp Assay ◽  
Trichuris Trichiura ◽  
Isothermal Amplification ◽  
Sub Saharan Africa ◽  
Health Concern ◽  
Alkaline Lysis ◽  
Loop Mediated Isothermal Amplification ◽  
Faecal Samples

Abstract Whipworm infection or trichuriasis caused by Trichuris trichiura is of major public health concern in sub-Saharan Africa, particularly among pre-school and school-going children. It is among the neglected tropical diseases targeted for elimination through mass drug administration (MDA). One of the outcomes of MDA is a rapid decline in levels of infection intensity, making it difficult to monitor effectiveness of control measures using the conventional Kato–Katz procedure, which relies on the microscopic detection of parasite ova in faecal samples. In the present study, a loop-mediated isothermal amplification (LAMP) test was developed for the detection of T. trichiura infection in faecal samples. LAMP technology offers greater sensitivity and specificity than the microscopy-based tests. A set of four specific primers targeting the internal transcribed spacer 2 region of the ribosomal DNA were designed using Primer Explorer software. DNA was extracted from faecal samples using the alkaline lysis method (HotSHOT) and the LAMP reaction performed at 63°C for 1 h. The amplicons were visualized by both gel electrophoresis and with the naked eye following staining with SYBR green dye. Sensitivity and specificity tests were determined using the standard Kato–Katz diagnostic procedure as a reference test. The developed LAMP assay reliably detected T. trichiura DNA in faecal samples, with a specificity and sensitivity of 88% and 77%, respectively. No cross-reactivity was observed with several common helminth parasites. The developed LAMP assay is an appropriate diagnostic method for the detection of T. trichiura DNA in human faecal samples due to its simplicity, low cost, high sensitivity and specificity.

scholarly journals Evaluation of Loop-Mediated Isothermal Amplification (LAMP) for Malaria Diagnosis in a Field Setting

2011 ◽  
Vol 85 (4) ◽  
pp. 594-596 ◽  
Author(s):  
Jeeraphat Sirichaisinthop ◽  
Risa Watanabe ◽  
Sureemas Buates ◽  
Takafumi Tsuboi ◽  
Satoru Takeo ◽  
...  
Keyword(s):  
Malaria Diagnosis ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Field Setting

scholarly journals High diagnostic performance of a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP) kit for the rapid detection of SARS-CoV-2

2021 ◽  
Author(s):  
Alexander Domnich ◽  
Andrea Orsi ◽  
Donatella Panatto ◽  
Vanessa De Pace ◽  
Valentina Ricucci ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Diagnostic Performance ◽  
Point Of Care ◽  
Lamp Assay ◽  
Laboratory Diagnosis ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Rt Pcr ◽  
Laboratory Equipment ◽  
Loop Mediated Isothermal Amplification

Abstract Although the reverse transcription polymerase chain reaction (RT-PCR) is considered a standard-of-care assay for the laboratory diagnosis of SARS-CoV-2, several limitations of this method have been described. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an alternative molecular assay and is potentially able to overcome some intrinsic shortcomings of RT-PCR. In this study, we evaluated the diagnostic performance of the novel HG COVID-19 RT-LAMP assay. In this retrospective analysis, a total of 400 routinely collected leftover nasopharyngeal samples with a known RT-PCR result were tested by means of the HG COVID-19 RT-LAMP assay. The overall sensitivity and specificity values of HG COVID-19 RT-LAMP versus RT-PCR were 97.0% (95% CI: 93.6–98.9%) and 98.5% (95% CI: 95.7–99.7%), respectively. Inter-assay agreement was almost perfect (κ = 0.96). Concordance was perfect in samples with high viral loads (cycle threshold <30). The average time to a positive result on RT-LAMP was 17 min. HG COVID-19 RT-LAMP is a reliable molecular diagnostic kit for detecting SARS-CoV-2, and its performance is comparable to that of RT-PCR. Shorter turnaround times and the possibility of performing molecular diagnostics in the point-of-care setting make it a valuable option for facilities without sophisticated laboratory equipment.

scholarly journals Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Somayyeh Sedaghatjoo ◽  
Monika K. Forster ◽  
Ludwig Niessen ◽  
Petr Karlovsky ◽  
Berta Killermann ◽  
...  
Keyword(s):  
Test Performance ◽  
Genomic Dna ◽  
Lamp Assay ◽  
Fungal Species ◽  
Isothermal Amplification ◽  
Genome Comparison ◽  
Performance Study ◽  
Loop Mediated Isothermal Amplification ◽  
Tilletia Controversa ◽  
Genomic Regions

AbstractTilletia controversa causing dwarf bunt of wheat is a quarantine pathogen in several countries. Therefore, its specific detection is of great phytosanitary importance. Genomic regions routinely used for phylogenetic inferences lack suitable polymorphisms for the development of species-specific markers. We therefore compared 21 genomes of six Tilletia species to identify DNA regions that were unique and conserved in all T. controversa isolates and had no or limited homology to other Tilletia species. A loop-mediated isothermal amplification (LAMP) assay for T. controversa was developed based on one of these DNA regions. The specificity of the assay was verified using 223 fungal samples comprising 43 fungal species including 11 Tilletia species, in particular 39 specimens of T. controversa, 92 of T. caries and 40 of T. laevis, respectively. The assay specifically amplified genomic DNA of T. controversa from pure cultures and teliospores. Only Tilletia trabutii generated false positive signals. The detection limit of the LAMP assay was 5 pg of genomic DNA per reaction. A test performance study that included five laboratories in Germany resulted in 100% sensitivity and 97.7% specificity of the assay. Genomic regions, specific to common bunt (Tilletia caries and Tilletia laevis together) are also provided.

Sign in / Sign up

Export Citation Format

Share Document