Application of a low-cost, specific, and sensitive loop-mediated isothermal amplification (LAMP) assay to detect Plasmodium falciparum imported from Africa

Zhang, Jiaqi; Chen, Xi; Pan, Maohua; Qin, Yucheng; Zhao, Hui; Yang, Qi; Li, Xinxin; Zeng, Weilin; Xiang, Zheng; Wu, Yuankui; Duan, Mengxi; Li, Xiaosong; Wang, Xun; Mazier, Dominique; Zhang, Yanmei; Zhu, Wei; Sun, Kemin; Wu, Yanhong; Huang, Yaming; Yang, Zhaoqing

doi:10.1016/j.molbiopara.2022.111529

Cited by 3 publications

(3 citation statements)

References 55 publications

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“…Actually, the plasmid DNA sample is a mixture of supercoiled, nick circular, and linearized forms. Following up with the same strategy of Chih-Hui Lin’s and others’ papers [ 24 , 25 , 26 , 27 , 28 , 29 ], in our present study, we directly quantified (averaged by five measurements) the plasmid DNA sample without enzyme treatment to make a rough and reasonable estimation. We think this may reduce concentration estimation error to some extent by averaging different conformation forms and does not significantly impact the conclusions of our study.…”

Section: Methodsmentioning

confidence: 99%

A Rapid and Sensitive Detection of HIV-1 with a One-Pot Two-Stage Reverse Transcription Recombinase Aided Real-Time PCR Assay

Tian

Jin

et al. 2023

TropicalMed

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Human immunodeficiency virus 1 (HIV-1) attacks the immune system, making people susceptible to various diseases, thus increasing their risk of death. Comprehensive detection of major HIV-1 strains circulating in China is vital for effective HIV-1 infection prevention and treatment. HIV-1 nucleic acid detection is considered effective for HIV-1 diagnosis since traditional immunological testing may fail to detect HIV-1 infection during the window period. This work demonstrates a one-pot two-stage amplification assay (RT-RAP), a combination of reverse transcription recombinase (RT- RAA), and quantitative real-time polymerase chain reaction (qRT-PCR). The turn-around time of the assay is only 50 min and can be performed with commonly available laboratory equipment, the qPCR devices. The RT-RAP assay could detect approximately 5 and 14 copies/reaction of HIV-1 DNA and RNA using recombinant plasmids and standard reference strains, respectively. Additionally, we found that the clinical performance of RT-RAP (detected 169 samples out of 170 specimens) was consistent with that of qRT-PCR. The sensitivity and specificity of RT-RAP were 100.00% (99/99) and 98.59% (70/71), respectively, while its positive and negative predictive values were 99.00% (99/100) and 100.00% (70/70), respectively. The total coincidence rate of the RT-RAP was 99.41% (169/170), with a kappa value of 0.988 (p < 0.05). We demonstrated that RT-RAP could rapidly detect the common HIV-1 subtypes commonly circulating in China with comparable sensitivity and specificity to qRT-PCR.

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Section: Methodsmentioning

confidence: 99%

A Rapid and Sensitive Detection of HIV-1 with a One-Pot Two-Stage Reverse Transcription Recombinase Aided Real-Time PCR Assay

Tian

Jin

et al. 2023

TropicalMed

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“…In addition, LAMP results can be interpreted using either colorimetic and fluorescent or Lateral flow detection [65]. Detailed procedures and protocol of LAMP technique for malaria diagnosis heavily depend on the specific type of kit used, as there are different commercially available LAMP kits and each with its specifications, for example, Illumigene (Alethia), malaria LAMP, and Loopamp malaria amplification kit [66]. One of the short-comings of this technique it is qualitative, not quantitative, but has a very good sensitivity similar to other sensitive molecular techniques like PCR [67].…”

Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

Advanced Techniques and Unusual Samples for Malaria Diagnosis

Muhammad,

Pukuma Sale,

Mahmoud Mohammed

2024

Infectious Diseases

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Successful malaria control, treatment, and prevention depends on successful diagnosis using appropriate equipment with high sensitivity and specificity. In most tropical countries where the disease is endemic, malaria diagnosis is still based on the conventional techniques (Microscopy and RDT) which have so many shortcomings, hence the need to switch to the most advanced diagnostic technique for better results. In this review, several serological and molecular malaria diagnostic techniques like Polymerase Chain Reaction (PCR), Flow cytometry, Loop-mediated Isothermal Amplification (LAMP), Indirect Immunofluorescence, Enzyme-Linked Immunosorbent Assay (ELISA), Radioimmunoassay (RIA), Quantitative Buffy Coat (QBC) and Laser Desorption Mass Spectrometry (LDMS) were systematically discussed in simple and direct language for easier understanding of the principle involved in each case scenario. In addition, some unusual samples for malaria diagnosis like Urine and saliva were also discussed.

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“…At present, PCR-based methods such as nested and real-time PCR are widely used at reference laboratories providing high accuracy and sensitivity and can diagnose mixed infections [ 9 ]. However, these methods are costly, requiring sophisticated equipment and professional technicians, making them difficult to use in many regions where malaria is endemic.…”

Section: Introductionmentioning

confidence: 99%

First field and laboratory evaluation of LAMP assay for malaria diagnosis in Cubal, Angola

Febrer-Sendra,

Crego-Vicente,

Nindia

et al. 2023

Parasites Vectors

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Background Malaria is a globally distributed infectious disease. According to the World Health Organization, Angola is one of the six countries that account for over half the global malaria burden in terms of both malaria cases and deaths. Diagnosis of malaria still depends on microscopic examination of thin and thick blood smears and rapid diagnostic tests (RDTs), which often lack analytical and clinical sensitivity. Molecular methods could overcome these disadvantages. The aim of this study was to evaluate, for the first time to our knowledge, the performance of a loop-mediated isothermal amplification (LAMP) for the diagnosis of malaria in an endemic area in Cubal, Angola, and to assess the reproducibility at a reference laboratory. Methods A total of 200 blood samples from patients attended at Hospital Nossa Senhora da Paz, Cubal, Angola, were analysed for Plasmodium spp. detection by microscopy, RDTs, and LAMP. LAMP assay was easily performed in a portable heating block, and the results were visualized by a simple colour change. Subsequently, the samples were sent to a reference laboratory in Spain to be reanalysed by the same colorimetric LAMP assay and also in real-time LAMP format. Results In field tests, a total of 67/200 (33.5%) blood samples were microscopy-positive for Plasmodium spp., 98/200 RDT positive, and 112/200 (56%) LAMP positive. Using microscopy as reference standard, field LAMP detected more microscopy-positive samples than RDTs (66/67; 98% vs. 62/67; 92.5%). When samples were reanalysed at a reference laboratory in Spain using both colorimetric and real-time assays, the overall reproducibility achieved 84.5%. Conclusions This is the first study to our knowledge in which LAMP has been clinically evaluated on blood samples in a resource-poor malaria-endemic area. The colorimetric LAMP proved to be more sensitive than microscopy and RDTs for malaria diagnosis in field conditions. Furthermore, LAMP showed an acceptable level of reproducibility in a reference laboratory. The possibility to use LAMP in a real-time format in a portable device reinforces the reliability of the assay for molecular diagnosis of malaria in resource-poor laboratories in endemic areas. Graphical Abstract

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Application of a low-cost, specific, and sensitive loop-mediated isothermal amplification (LAMP) assay to detect Plasmodium falciparum imported from Africa

Cited by 3 publications

References 55 publications

A Rapid and Sensitive Detection of HIV-1 with a One-Pot Two-Stage Reverse Transcription Recombinase Aided Real-Time PCR Assay

A Rapid and Sensitive Detection of HIV-1 with a One-Pot Two-Stage Reverse Transcription Recombinase Aided Real-Time PCR Assay

Advanced Techniques and Unusual Samples for Malaria Diagnosis

First field and laboratory evaluation of LAMP assay for malaria diagnosis in Cubal, Angola

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