Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time
Kayla G. BarnesAnna E. LachenauerAdam D. NitidoSameed M. SiddiquiRobin GrossBrett BeitzelKatherine J. SiddleCatherine A. FreijeBonnie Dighero-KempSamar MehtaAmber CarterJessica N. UwanibeFehintola V. AjogbasileTestimony OlumadeIkponmwosa OdiaJohn Demby SandiMambu MomohHayden C. MetskyChloe K. BoehmAaron E. LinMolly KemballDaniel J. ParkLuis M. BrancoMatthew L. BoisenBrian M. SullivanMihret F. AmareAbdulwasiu Bolaji TiamiyuZahra ParkerMichael IroezinduDonald S. GrantKayvon ModjarradCameron MyhrvoldRobert F. GarryGustavo PalaciosLisa E. HensleyS. F. SchaffnerChristian T. HappiAndrés ColubriPardis C. Sabeti
131
Citation
32
Reference
10
Related Paper
Citation Trend
Abstract:
Abstract Recent outbreaks of viral hemorrhagic fevers (VHFs), including Ebola virus disease (EVD) and Lassa fever (LF), highlight the urgent need for sensitive, deployable tests to diagnose these devastating human diseases. Here we develop CRISPR-Cas13a-based (SHERLOCK) diagnostics targeting Ebola virus (EBOV) and Lassa virus (LASV), with both fluorescent and lateral flow readouts. We demonstrate on laboratory and clinical samples the sensitivity of these assays and the capacity of the SHERLOCK platform to handle virus-specific diagnostic challenges. We perform safety testing to demonstrate the efficacy of our HUDSON protocol in heat-inactivating VHF viruses before SHERLOCK testing, eliminating the need for an extraction. We develop a user-friendly protocol and mobile application (HandLens) to report results, facilitating SHERLOCK’s use in endemic regions. Finally, we successfully deploy our tests in Sierra Leone and Nigeria in response to recent outbreaks.Keywords:
Lassa virus
Lassa fever
Viral hemorrhagic fever
Lassa virus, the causative agent of Lassa fever, is a zoonotic pathogen causing annual outbreaks in West African countries. Human patients can develop lethal hemorrhagic fever in severe cases. Although Lassa virus is one of the most alarming pathogens from a public health perspective, there are few available countermeasures, such as antiviral drugs or vaccines. Moreover, the fact that animal models are not readily accessible and the fact that mostly laboratory viruses, which have been passaged many times after isolation, are used for studies further limits the successful development of countermeasures. In this study, we demonstrate that a human isolate of Lassa virus causes lethal infection uniformly in Hartley guinea pigs. This novel animal model of Lassa fever may contribute to Lassa fever research and the development of vaccines and therapeutics.
Lassa virus
Lassa fever
New guinea
Cite
Citations (14)
Lassa fever
Arenavirus
Lassa virus
Viral hemorrhagic fever
Mastomys
Cite
Citations (0)
Abstract Recent outbreaks of viral hemorrhagic fevers (VHFs), including Ebola virus disease (EVD) and Lassa fever (LF), highlight the urgent need for sensitive, deployable tests to diagnose these devastating human diseases. Here we develop CRISPR-Cas13a-based (SHERLOCK) diagnostics targeting Ebola virus (EBOV) and Lassa virus (LASV), with both fluorescent and lateral flow readouts. We demonstrate on laboratory and clinical samples the sensitivity of these assays and the capacity of the SHERLOCK platform to handle virus-specific diagnostic challenges. We perform safety testing to demonstrate the efficacy of our HUDSON protocol in heat-inactivating VHF viruses before SHERLOCK testing, eliminating the need for an extraction. We develop a user-friendly protocol and mobile application (HandLens) to report results, facilitating SHERLOCK’s use in endemic regions. Finally, we successfully deploy our tests in Sierra Leone and Nigeria in response to recent outbreaks.
Lassa virus
Lassa fever
Viral hemorrhagic fever
Cite
Citations (131)
Abstract Lassa fever is a neglected tropical disease with a significant impact on the health care system of endemic West African nations. To date, case reports of Lassa fever have focused on laboratory characterisation of serological, biochemical and molecular aspects of the disease imported by infected individuals from Western Africa to the United States, Canada, Europe, Japan and Israel. Our report presents the first comprehensive real time diagnosis and characterization of a severe, hemorrhagic Lassa fever case in a Sierra Leonean individual admitted to the Kenema Government Hospital Lassa Fever Ward. Fever, malaise, unresponsiveness to anti-malarial and antibiotic drugs, followed by worsening symptoms and onset of haemorrhaging prompted medical officials to suspect Lassa fever. A recombinant Lassa virus protein based diagnostic was employed in diagnosing Lassa fever upon admission. This patient experienced a severe case of Lassa hemorrhagic fever with dysregulation of overall homeostasis, significant liver and renal system involvement, the interplay of pro- and anti-inflammatory cytokines during the course of hospitalization and an eventual successful outcome. These studies provide new insights into the pathophysiology and management of this viral illness and outline the improved infrastructure, research and real-time diagnostic capabilities within LASV endemic areas.
Lassa fever
Lassa virus
Viral hemorrhagic fever
Arenavirus
Malaise
Cite
Citations (44)
The peace agreement signed last month, which will see the demo- cratically elected president back in power in April, 1998, should have been good news for Sierra Leone. But the country is still in the grip of the worst-ever Lassa-fever epidemic. And continued fighting is making conditions increasingly difficult.
Lassa fever
Lassa virus
Cite
Citations (0)
Introduction: viral hemorrhagic fever is a generic term for a severe illness, often accompanied by bleeding and caused by viruses. Lassa fever is a viral hemorrhagic fever caused by Lassa Virus of the family Arenaviridae. Lassa fever is endemic in Nigeria especially Edo State with zero-prevalence of 21% and 28% mortality. The aim of this study was to evaluate the laboratory based surveillance system for Lassa fever to assess its key attributes and whether it meets its objectives.
Lassa fever
Lassa virus
Arenavirus
Viral hemorrhagic fever
Cite
Citations (2)
Lassa fever is a viral hemorrhagic fever caused by Lassa virus. The Lassa virus is an enveloped single-stranded, non-lytic bi-segmented negative-stranded RNA virus belonging to the family Arenaviridae. Lassa fever is relatively common or endemic in West Africa or particularly in countries such as Benin, Ghana, Guinea, Liberia, Mali, Sierra Leone, and Nigeria. Humans usually become infected with the virus through exposure to food or household items contaminated with the urine or faeces of infected multimammate rat while spread of the disease between or among persons is by direct contact. The disease cases rises to the peak during the dry season particularly between December to April and falls in May annually. The incubation period for the disease is between 1 – 3 weeks, which can lead to spread from region to region. This review focuses on the secondary data for the epidemiological trend of the Lassa fever disease in Nigeria. The rodent host and reservoir is the primary driver of the Lassa fever seasonal trends. Therefore, to control the disease, it is necessary to control the rodent host by killing it. This can be achieved through the use of trap,poisoned bait and interference with their breeding by killing the newly born off springs.
Lassa fever
Lassa virus
Arenavirus
Viral hemorrhagic fever
Mastomys
Cite
Citations (1)
Viral hemorrhagic fever (VHF) is defined as virus infections that usually cause pyrexia and hemorrhagic symptoms with multiple organ failure. VHF includes following viral infections: Ebola hemorrhagic fever (EHF), Marburg hemorrhagic fever (MHF), Crimean-Congo hemorrhagic fever (CCHF) and Lassa fever. In particular, the causative agents of EHF, MHF, CCHF, and Lassa fever are Ebola, Marburg, CCHF, Lassa viruses, respectively, and regarded as biosafety level-4 pathogens because of their high virulence to humans. Recently, relatively large outbreaks of EHF and MHF have occurred in Africa, and areas of EHF- and MHF-outbreaks seem to be expanding. Although outbreaks of VHF have not been reported in Japan, there is a possibility that the deadly hemorrhagic fever viruses would be introduced to Japan in future. Therefore, preparedness for possible future outbreaks of VHF is necessary in areas without VHF outbreaks.
Viral hemorrhagic fever
Lassa fever
Lassa virus
Crimean–Congo hemorrhagic fever
Ebola Hemorrhagic Fever
Marburg virus
Filoviridae
Preparedness
Cite
Citations (1)
Lassa fever
Lassa virus
Viral hemorrhagic fever
Cite
Citations (0)
Background. Kenema Government Hospital (KGH) has developed an advanced clinical and laboratory research capacity to manage the threat of Lassa fever, a viral hemorrhagic fever (VHF). The 2013–2016 Ebola virus (EBOV) disease (EVD) outbreak is the first to have occurred in an area close to a facility with established clinical and laboratory capacity for study of VHFs. Methods. Because of its proximity to the epicenter of the EVD outbreak, which began in Guinea in March 2014, the KGH Lassa fever Team mobilized to establish EBOV surveillance and diagnostic capabilities. Results. Augustine Goba, director of the KGH Lassa laboratory, diagnosed the first documented case of EVD in Sierra Leone, on 25 May 2014. Thereafter, KGH received and cared for numbers of patients with EVD that quickly overwhelmed the capacity for safe management. Numerous healthcare workers contracted and lost their lives to EVD. The vast majority of subsequent EVD cases in West Africa can be traced back to a single transmission chain that includes this first diagnosed case. Conclusions. Responding to the challenges of confronting 2 hemorrhagic fever viruses will require continued investments in the development of countermeasures (vaccines, therapeutic agents, and diagnostic assays), infrastructure, and human resources.
Lassa fever
Viral hemorrhagic fever
Lassa virus
Ebolavirus
Ebola Hemorrhagic Fever
Cite
Citations (36)