Rapid Detection of Zika Virus in Urine Samples and Infected Mosquitos by Reverse Transcription-Loop-Mediated Isothermal Amplification

Lamb, Laura E.; Bartolone, Sarah N.; Tree, Maya O.; Conway, Michael J.; Rossignol, Julien; Smith, Christopher P.; Chancellor, Michael B.

doi:10.1038/s41598-018-22102-5

Cited by 54 publications

(66 citation statements)

References 26 publications

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“…Previously published papers demonstrated the efficacy of RT-LAMP compared with RT-PCR to detect ZIKV [18–20]. The LAMP assay has shown to be useful to detect the ZIKV RNA in saliva and urine samples.…”

Section: Discussionmentioning

confidence: 99%

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Castro

Sabalza

Barber

et al. 2018

Journal of Oral Microbiology

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Background: Zika virus (ZIKV) is a single-stranded RNA virus and member of the Flaviviridae family. Recent studies have reported that saliva can be an important alternative to detect ZIKV. Saliva requires less processing than blood greatly simplifying the assay. Loop-mediated Isothermal Amplification (LAMP) is a rapid assay that detects nucleic acids, including ZIKV RNA. Aim: The aim of this study was to evaluate the efficacy of saliva and urine to diagnose ZIKV infection in subjects during the acute phase, through ZIKV RNA detection by LAMP. Method: A total of 131 samples (68 saliva and 63 urine samples) from 69 subjects in the acute phase of ZIKV infection, and confirmed positive for ZIKV by blood analysis through real time-PCR, were collected and analyzed by Reverse Transcriptase Loop-mediated Isothermal Amplification (RT-LAMP). Results: From the 68 saliva samples, 45 (66.2%) were positive for ZIKV with an average time to positivity (Tp) of 13.5 min, and from the 63 urine samples, 25 (39.7%) were positive with the average Tp of 15.8 min. Saliva detected more samples (p = 0.0042) and had faster Tp (p = 0.0176) as compared with urine. Conclusion: Saliva proved to be a feasible alternative to diagnose ZIKV infection during the acute phase by LAMP.

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Section: Discussionmentioning

confidence: 99%

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Castro

Sabalza

Barber

et al. 2018

Journal of Oral Microbiology

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“…Some highlights from adaptations of LAMP include multiplexed LAMP, electrochemical LAMP, and disc-based LAMP [46][47][48]. For RNA viruses, such as ZIKV, DENV, and CHIKV, it is necessary to perform a reverse transcription reaction LAMP (RT-LAMP) [27,39,42,[49][50][51][52] which includes the enzymes that first convert RNA → DNA upstream of the LAMP process, unless a LAMP enzyme with both reverse transcriptase and DNA polymerase is used [53].…”

Section: Principles Of Lamp Assaymentioning

confidence: 99%

“…The LAMP assay can be performed using either a one-step or two-step protocol directly from sample matrices, including serum, semen, urine, saliva, and mosquitoes homogenate. The use of Bst 2.0 polymerase, Bst 3.0 polymerase or OmniAmp polymerase enables the test to be performed in a one-step protocol [38,[40][41][42]53,65] (Figure 2). These enzymes have both reverse transcriptase (RT) and DNA polymerase activities at a fixed temperature (50-72 °C), and so can be run isothermally for Abbreviations: aM, attomolar; PFU, plaque-forming unit; FFU, focus forming units; TCID 50 , 50% tissue culture infective dose.…”

Section: Step 1: Template Preparationmentioning

confidence: 99%

“…Importantly, the simple, single-temperature incubation allows LAMP reactions to be performed without expensive equipment, directly in the field [36]. Since the 2015 emergence of the ZIKV in Brazil, many LAMP assays have been developed for diagnosis by research groups across the world [27,[37][38][39][40][41][42][43][44][45]. These diagnostic platforms based on LAMP have proven to be specific, sensitive and inexpensive POC tools that can be applied even in resource-limited regions of the world.…”

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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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“…We have previously used this method to detect zika virus in clinical serum and urine samples as well as mosquitos. 4,5 This study describes a RT-LAMP methodology that can detect COVID-19 in simulated patient samples in under 30 minutes. The test could be used at the point-of-care by field and local personnel for the rapid diagnosis of individuals for optimal treatment, isolation, and rapid contact tracking as well as the investigation of outbreaks of unknown respiratory diseases.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Novel Coronavirus (COVID-19) by Reverse Transcription-Loop-Mediated Isothermal Amplification

Lamb

Bartolone

Ward

et al. 2020

Preprint

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Novel Corona virus (COVID-19 or 2019-nCoV) is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for COVID-19 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in under 30 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the COVID-19 nucleic sequence. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected COVID-19 in simulated patient samples.This test was performed in under 30 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

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Rapid Detection of Zika Virus in Urine Samples and Infected Mosquitos by Reverse Transcription-Loop-Mediated Isothermal Amplification

Cited by 54 publications

References 26 publications

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

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

Rapid Detection of Novel Coronavirus (COVID-19) by Reverse Transcription-Loop-Mediated Isothermal Amplification

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