Development and Evaluation of a One-Step Quantitative RT-PCR Assay for Detection of Lassa Virus

Dedkov, Vladimir G.; Magassouba, N’Faly; Safonova, Marina V.; Naydenova, E. V.; Ayginin, Andrey A.; Soropogui, Barré; Kourouma, Fodé; Camara, A.; Camara, Jacob; Kritzkiy, Andrey A.; Tuchkov, I. V.; Shchelkanov, M. Yu.; Maleev, V V

doi:10.1016/j.jviromet.2019.113674

Cited by 10 publications

(13 citation statements)

References 34 publications

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“…[24], и методом фиксации комплемента антителами сывороток крови реконвалесцентов [83]. Методы научных исследований LASV включают: анализы на основе полимеразной цепной реакции (ПЦР) по обнаружению вирусной РНК, электронную микроскопию, выделение инфекционного вируса на культуре чувствительных клеток, реакцию непрямой иммунофлуоресценции (РНИФ), иммуноферментный (ИФА) и иммунохроматографический (ИХА) анализы по выявлению антител и/или антигена, а также иммуноблоттинг [7,10,27,77]. Для диагностики ЛЛ в настоящее время используют, в основном, тест-системы на основе молекулярно-генетических методов [27,66].…”

Section: методы научных исследований Lasv и лабораторной диагностики ллunclassified

“…С появлением в базе данных GenBank [https://pubmed.ncbi.nlm.nih.gov/genbank] дополнительной информации о генетических последовательностях LASV начались исследования по разработке анализов ОТ-ПЦР, нацеленных на L-сегмент РНК, кодирующий полимеразу и белок Z [27,41]. За последние годы большое количество геномов LASV были полностью секвенированы и оказалось, что последовательности L-генов также демонстрируют высокую генетическую изменчивость, как и последовательности генов GPC и NP, даже в пределах одной генетической линии.…”

Section: молекулярно-генетические методы исследованияunclassified

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Lassa fever. Part 2. Laboratory diagnostics, treatment, development of medications

Казачинская

Aripov

Ivanova

et al. 2022

Russian Journal of Infection and Immunity

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Abstract. Globalization and high-speed means of transportation contribute to the spread of infections dangerous to humans. Airborne pathogens have pandemic potential as currently shown in case of the novel coronavirus SARS-CoV-2. Natural focal Lassa fever (LF) common in West African countries, in 35 cases was registered in non-endemic geographical areas because any person infected with Lassa virus (LASV) is a long-term source of infection (up to two months). Cases of person-to-person infection in endemic territories are described. In Germany, the facts of secondary virus transmission from patients to doctors have been recorded during the examination and blood collection from an apparently healthy person as well as during the autopsy of a deceased subjects due to severe LF course.Nonspecific malaise symptoms in LF are also characteristic of numerous other diseases common on the African continent, e.g., malaria and typhoid fever or viral infections such as yellow fever, Chikungunya, dengue and Zika, monkey pox and Ebola virus disease. In this regard, there may be similar dermatological manifestations. Timely detection of cases and differential diagnosis are crucial to ensure safe patient care and use of affordable antiviral therapy for LL provided by the drug ribavirin.research methods for studying LASV use polymerase chain reaction (PCR) for detecting viral RNA, electron microscopy, isolation of infectious virus cultured sensitive cells, indirect immunofluorescence reaction, enzyme immunoassay (ELISA) and immunochromatographic assays for the detection of antibodies and /or antigen as well as immunoblotting. Currently, test kits based on molecular and genetic methods are mainly used for LF laboratory diagnostics.Since the 1980s, ribavirin has been used to treat patients with LF. The serum accumulation of the drug in large quantities causes hemolysis, development of anemia and impaired renal function. In this regard, treatment options are being considered with decline in its concentration due to combined use with other antiviral drugs. A search for new therapeutic agents capable of inhibiting viral replication at disease early stage has been in progress due to lack of any approved vaccines.

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Section: методы научных исследований Lasv и лабораторной диагностики ллunclassified

Section: молекулярно-генетические методы исследованияunclassified

Lassa fever. Part 2. Laboratory diagnostics, treatment, development of medications

Казачинская

Aripov

Ivanova

et al. 2022

Russian Journal of Infection and Immunity

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“… Virus Sample source Analytical Method No. of samples Reference HBV, HCV serum PCR 730 [ 92 ] Enteric viruses stool samples monoplex or multiplex RT-PCR 227 [ 93 ] HRSV nasopharyngeal and oropharyngeal swab real-time RT-PCR 334 [ 94 ] DENV, ZIKV serum RT-qPCR 46 [ 95 ] Respiratory viruses throat swabs and sputum samples RT-qPCR 408 [ 96 ] Respiratory viruses nasopharyngeal aspirates, nasopharyngeal and oral pharyngeal swab rRT-PCR 8173 [ 97 ] SARS-CoV-2 sputum, nose and throat swabs real-time RT-PCR 297 [ 98 ] LASV serum, lung and spleen tissue, blood RT-qPCR 82 [ 99 ] Respiratory viruses respiratory tract swabs and aspirates MALDI-TOF MS 58 [ 89 ] HR HPV uterine cervix cytology MALDI-TOF MS 356 [ 90 ] Poliovirus cerebrospinal fluid, throat swab, stool samples MALDI-TOF MS 5 [ 88 ] SARS-CoV-2 nasal swab MALDI-MS + machine learning analysis 362 [ 39 ] Enterovirus EV71 serum …”

Section: Sample Preparation Techniques For Pathogen Determinationmentioning

confidence: 99%

A new approach to identifying pathogens, with particular regard to viruses, based on capillary electrophoresis and other analytical techniques

Buszewski

Maślak

Złoch

et al. 2021

TrAC Trends in Analytical Chemistry

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“…The PCR requirement for instrumentation and preextraction step of RNA increases the cost and TAT with the consequences of extending the mortality rate of patients [37]. The quantitative reverse transcriptase PCR (RT-qPCR) for the detection of LASV is highly crucial for improving POC diagnosis and for surveillance and epidemiological control studies [38]. Both SYBR-Green and TaqMan probe methods are readily applicable for LASV detection.…”

Section: Reverse Transcription Polymerase Chain Reaction (Rt-pcr) Plamentioning

confidence: 99%

“…The less specificity of the SYBR-Green-based method is associated with fluorescent dyes that intercalate into any formed double-stranded DNA including primer-dimers and non-specific amplified products [39]. The main advantage of RT-qPCR is that it allows verification of all prior steps of analysis including extraction, reverse transcription, and PCR amplification through melting and standard curve analyses [38]. The quantification capability of RT-qPCR also plays a significant role in strain identification.…”

Section: Reverse Transcription Polymerase Chain Reaction (Rt-pcr) Plamentioning

confidence: 99%

Lassa Virus Diagnostic Platforms: Limitations and Prospects

Hassan

Kumurya

Aminu

et al. 2020

AJRID

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Background: Lassa virus (LASV) is the cause of lassa fever (LF) belonging to the Arenaviridae family. Clinical diagnosis is often difficult because of symptoms commonality with other infectious diseases. Early and rapid diagnosis is critical for therapy initiation and LF transmission prevention and control. Aims: This review aims to highlight current diagnostic platforms and prospects of new emerging sensitive platforms. Methodology: Available published articles on LASV diagnostics with a focus on current methods: virus culture, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT – PCR) and rapid diagnostic tests (RDT) were reviewed based on their performances and limitations. Prospects of new diagnostic platforms: mobile health, microfluidic, clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas), Loop-mediated isothermal amplification (LAMP) for LASV diagnosis were also reviewed. Results: Low sensitivity of the ELISA platform during the window period of LASV infection was observed. Moreover, RT – PCR findings indicated limitation of expertise necessity, cost of thermal cycler, and dedicated facility. Molecular-based point-of-care (POC) diagnostic development should be prioritized to increase speed and sensitivity. Conclusion: The integration of POC device into molecular isothermal method against LASV scourge will be a success story in curving intermittent outbreaks in endemic areas and prompt clinical management.

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Development and Evaluation of a One-Step Quantitative RT-PCR Assay for Detection of Lassa Virus

Cited by 10 publications

References 34 publications

Lassa fever. Part 2. Laboratory diagnostics, treatment, development of medications

Lassa fever. Part 2. Laboratory diagnostics, treatment, development of medications

A new approach to identifying pathogens, with particular regard to viruses, based on capillary electrophoresis and other analytical techniques

Lassa Virus Diagnostic Platforms: Limitations and Prospects

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