Ten-minute direct detection of Zika virus in serum samples by RT-LAMP

Estrela, Paulo Felipe Neves; Mendes, Geovana de Melo; Oliveira, Kézia Gomes de; Bailão, Alexandre Melo; Soares, Célia Maria de Almeida; Assunção, Nilson Antônio; Duarte, Gabriela Rodrigues Mendes

doi:10.1016/j.jviromet.2019.113675

Cited by 27 publications

(26 citation statements)

References 24 publications

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“…In establishing a new LAMP diagnostic assay, it is important to optimize the protocol and include the proper controls to limit the risk of false positives or non-target amplification [40]. Variables to consider in LAMP optimization include Mg 2+ concentrations, dNTP mix concentrations, primers, time reaction, and temperature reaction, which all have a key role in producing reliable results [40,42,44,64]. Lower LAMP temperatures and excessive free Mg 2+ ions are associated with non-target amplification [27], and, indeed, previous studies have reported false-positive results in RT-LAMP assays [40,41,44,64].…”

Section: Step 2: Practical Implementationmentioning

confidence: 99%

“…Neves et al (2019) developed a simple system based on an RT-LAMP assay using Bst DNA polymerase 3.0 without RNA extraction or sample pre-treatment [64]. They reported that the reaction time was the most important factor to avoid non-target amplification.…”

Section: Step 2: Practical Implementationmentioning

confidence: 99%

“…They reported that the reaction time was the most important factor to avoid non-target amplification. Evaluating four reaction durations (5,10,15, and 20 min), they found that 10-min incubations were sufficient to detect ZIKV from serum samples, whereas 20-min incubations resulted in false-positive amplifications in non-template control (NTC) using water [64].…”

Section: Step 2: Practical Implementationmentioning

confidence: 99%

“…Below we review each of these output modes, beginning with the fluorescent dyes calcein and SYBR green I dyes, which are some of the most widely used for the detection of LAMP amplicons. Here as target sequences are amplified; the dye binds becoming fluorescent and creating an optical readout of LAMP-mediated detection [42,53,64]. Although it presents adequate sensitivity, SYBR green I could inhibit LAMP reaction if the dye is added during the set-up of the LAMP reaction mixture, and when added after incubation, there is a real risk of cross-contamination between LAMP reaction tubes [68].…”

Section: Step 3: Lamp Modes Of Outputmentioning

confidence: 99%

“…Here positive reactions generate ladder-like patterns, which correspond to the series of inverted repeats of the amplified target gene and cauliflower-like structures obtained from LAMP products reactions. This has been a popular method of detection for LAMP-based ZIKV assays [27,39,40,42,44,53,64]. However, visualization of LAMP amplicons by gel electrophoresis is time-consuming and carries an increased risk of cross-contamination between samples due to the massive generation of DNA amplicons during the LAMP assay [46,84].…”

Section: Step 3: Lamp Modes Of Outputmentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Silva

Pardee

Pena

2019

Viruses

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The recent outbreak of Zika virus (ZIKV) in the Americas and its devastating developmental and neurological manifestations has prompted the development of field-based diagnostics that are rapid, reliable, handheld, specific, sensitive, and inexpensive. The gold standard molecular method for lab-based diagnosis of ZIKV, from either patient samples or insect vectors, is reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The method, however, is costly and requires lab-based equipment and expertise, which severely limits its use as a point-of-care (POC) tool in resource-poor settings. Moreover, given the lack of antivirals or approved vaccines for ZIKV infection, a POC diagnostic test is urgently needed for the early detection of new outbreaks and to adequately manage patients. Loop-mediated isothermal amplification (LAMP) is a compelling alternative to RT-qPCR for ZIKV and other arboviruses. This low-cost molecular system can be freeze-dried for distribution and exhibits high specificity, sensitivity, and efficiency. A growing body of evidence suggests that LAMP assays can provide greater accessibility to much-needed diagnostics for ZIKV infections, especially in developing countries where the ZIKV is now endemic. This review summarizes the different LAMP methods that have been developed for the virus and summarizes their features, advantages, and limitations.Viruses 2020, 12, 19 2 of 20 raising concerns about the neurological tropism of the virus [12]. In May 2015, the first case of ZIKV infection was reported in Brazil [13] and the virus rapidly spread throughout the country and much of Latin America, causing the largest recorded epidemic of the virus to date [14]. The Brazilian epidemic raised great international concern because of severe birth defects, including microcephaly, in neonates born to mothers infected by ZIKV during pregnancy [3,15,16].ZIKV is transmitted mainly through the bite of infected mosquitoes from the genus Aedes, although other vectors may also be involved in the transmission [17][18][19][20][21]. Additionally, other routes of ZIKV transmission have been identified, including blood transfusions, transplacental, perinatal, and sexual intercourse [22][23][24]. ZIKV infection usually causes a self-limiting and a mild illness, where the majority of cases are asymptomatic and, when present, symptoms include fever, headache, rash, conjunctivitis, and arthralgia [25]. In regions where there is a circulation of other arboviruses, such as DENV and chikungunya (CHIKV), the clinical diagnosis of ZIKV infection becomes extremely difficult because of common symptoms. Therefore, laboratory-based molecular diagnosis is of fundamental importance to correctly identify the etiologic agent [3,26].Given the lack of approved vaccines and antivirals against ZIKV, a rapid and reliable point-of-care (POC) diagnostic test for detection of ZIKV is urgently required for control and prevention measures and to increase the diagnostic capacity of ZIKV-affected, mainly in low-resource areas [27,28]. Z...

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Section: Step 2: Practical Implementationmentioning

confidence: 99%

Section: Step 2: Practical Implementationmentioning

confidence: 99%

Section: Step 2: Practical Implementationmentioning

confidence: 99%

Section: Step 3: Lamp Modes Of Outputmentioning

confidence: 99%

Section: Step 3: Lamp Modes Of Outputmentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Silva

Pardee

Pena

2019

Viruses

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Clinical and laboratory diagnosis of Mayaro virus (MAYV): Current status and opportunities for further development

da Silva,

Krokovsky

2024

Reviews in Medical Virology

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The recent outbreaks related to Mayaro virus (MAYV) infection in the Americas have brought this neglected virus as a potential threat to global public health. Given the range of symptoms that can be associated with MAYV infection, it can be challenging to diagnose individuals based on clinical signs, especially in countries with simultaneous circulation of other mosquito‐borne viruses, such as dengue virus (DENV) and chikungunya virus (CHIKV). With this challenge in mind, laboratory‐based diagnosis assumes a critical role in the introduction of measures to help prevent virus dissemination and to adequately treat patients. In this review, we provide an overview of the clinical features reported in infected patients and currently available laboratory tools that are used for MAYV diagnosis, discussing their advances, advantages, and limitations to apply in the field. Moreover, we explore novel point‐of‐care (PoC) diagnostic platforms that can provide de‐centralised diagnostics for use in areas with limited laboratory infrastructure.

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Validation of Zika virus infections: A focus on molecular methods

Jääskeläinen

2021

Zika Virus Biology, Transmission, and Pathology

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Ten-minute direct detection of Zika virus in serum samples by RT-LAMP

Cited by 27 publications

References 24 publications

Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Clinical and laboratory diagnosis of Mayaro virus (MAYV): Current status and opportunities for further development

Validation of Zika virus infections: A focus on molecular methods

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