Comparison of Fluorescent Intercalating Dyes for Quantitative Loop-Mediated Isothermal Amplification (qLAMP)

Oscorbin, Igor P.; Belousova, E. A.; Zakabunin, A. I.; Boyarskikh, U. A.; Филипенко, М. Л.

doi:10.2144/000114432

Cited by 96 publications

(74 citation statements)

References 25 publications

(17 reference statements)

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“…More recently, several research groups have reported the use of other dyes for end-point reads of LAMP assays, with potential applications for ZIKV. These include malachite green dye [72,73], hydroxynaphthol blue dye [74,75], evagreen dye [76,77], Goldview II dye [78], Gelred dye [79,80], SYTO fluorescent dye [81,82], and berberine dye [83].…”

Section: Step 3: Lamp Modes Of Outputmentioning

confidence: 99%

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 3: Lamp Modes Of Outputmentioning

confidence: 99%

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

Silva

Pardee

Pena

2019

Viruses

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“…Because SYBR Green I dye has several limitations, including inhibition of PCR, preferential binding to CG-rich sequences and effects on MCA, two intercalating dyes SYTO-13 and SYTO-82 were tried and did not show these negative effects, and SYTO-82 demonstrated a 50-fold lower detection limit (64), as well as best combinations of time-to threshold (Tt) and signal-to-noise ratio (SNR) (65). To optimize performance of the buffer, a PCR mix supplemented with two additives, 1M 1,2-propanediol and 0.2 M trehalose, were shown to decrease T m , efficiently neutralize PCR inhibitors, and increase the robustness and performance of qPCR with short amplicons (66).…”

Section: Melting Curve Analysis (Mca)mentioning

confidence: 99%

Role of Melt Curve Analysis in Interpretation of Nutrigenomics’ MicroRNA Expression Data

Ahmed

Gouda

Hussein

et al. 2017

CGP

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“…Well optimized LAMP assay shows sensitivity comparable to that of PCR, less than 10 copies per reaction [12]. Intercalating fluorescent dyes are compatible with LAMP reaction so that we can observe amplification in real-time [13]. Because amplification efficiency of LAMP reaction is very high, changes in reaction mixture components made it possible to detect the result with colorimetric detection methods [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Development of Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assays Targeting SARS-CoV-2

Park

Beak

et al. 2020

Preprint

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AbstractEpidemics of Coronavirus Disease 2019 (COVID-19) now have more than 100,000 confirmed cases worldwide. Diagnosis of COVID-19 is currently performed by RT-qPCR methods, but the capacity of RT-qPCR methods is limited by its requirement of high-level facilities and instruments. Here, we developed and evaluated RT-LAMP assays to detect genomic RNA of SARS-CoV-2, the causative virus of COVID-19. RT-LAMP assays in this study can detect as low as 100 copies of SARS-CoV-2 RNA. Cross-reactivity of RT-LAMP assays to other human Coronaviruses was not observed. We also adapted a colorimetric detection method for our RT-LAMP assay so that the tests potentially performed in higher throughput.

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Comparison of Fluorescent Intercalating Dyes for Quantitative Loop-Mediated Isothermal Amplification (qLAMP)

Cited by 96 publications

References 25 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

Role of Melt Curve Analysis in Interpretation of Nutrigenomics’ MicroRNA Expression Data

Development of Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assays Targeting SARS-CoV-2

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