OBJECTIVEMarburg virus and Ebola virus are acute infections with high case fatality rates. A rapid, sensitive detection method was established to detect Marburg virus and Ebola virus by multiplex real-time fluorescence quantitative PCR.

METHODSDesigning primers and Taqman probes from highly conserved sequences of Marburg virus and Ebola virus through whole genome sequences alignment, Taqman probes labeled by FAM and Texas Red, the sensitivity of the multiplex real-time quantitative PCR assay was optimized by evaluating the different concentrations of primers and Probes.

RESULTSWe have developed a real-time PCR method with the sensitivity of 30.5 copies/microl for Marburg virus positive plasmid and 28.6 copies/microl for Ebola virus positive plasmids, Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity.

CONCLUSIONSThe Multiplex real-time PCR assays provide a sensitive, reliable and efficient method to detect Marburg virus and Ebola virus simultaneously.

Ebolavirus ; genetics ; isolation & purification ; Filoviridae Infections ; diagnosis ; virology ; Hemorrhagic Fever, Ebola ; diagnosis ; virology ; Humans ; Marburgvirus ; genetics ; isolation & purification ; Multiplex Polymerase Chain Reaction ; methods ; Real-Time Polymerase Chain Reaction ; methods

Hemorrhagic Fever, Ebola ; pathology ; Humans

Ebola virus (EBOV) causes hemorrhagic fever, resulting in mortality rates as high as 90% among infected humans and non-human primates (NHPs). The 2014 Ebola epidemic in West Africa is the severest in history, leading to WHO taking all control measures to stop any possibility of cross-border outbreaks. Because no licensed vaccines or effective therapeutics against EBOV are available, the current outbreak management has been limited to palliative care and barrier methods to prevent transmission. Several promising experimental EBOV vaccines have demonstrated protection in NHPs against lethal EBOV challenge, and some progresses have been made through clinical trials of EBOV vaccine candidates. It is believed there will be some licensed vaccine available in the near future to control EBOV outbreaks. In this review we provide some insights for further development of EBOV vaccines.
Animals ; Ebola Vaccines ; Ebolavirus ; Hemorrhagic Fever, Ebola ; prevention & control ; Humans

Ebolavirus ; pathogenicity ; Endemic Diseases ; Hemorrhagic Fever, Ebola ; epidemiology ; virology ; Humans

Hemorrhagic Fever, Ebola ; diagnosis ; drug therapy ; epidemiology ; prevention & control ; Humans

Asia ; Disease Outbreaks ; Hemorrhagic Fever, Ebola ; epidemiology ; Humans

Ebola virus (EBOV) is an extremely contagious pathogen first discovered in Africa associated with severe hemorrhagic disease in humans and nonhuman primates, which has resulted in at least 28 500 suspected cases and 11 300 confirmed deaths in 2014-2016 Ebola epidemic in West Africa. Rapid and sensitive detection of EBOV is the key to increasing the probability of survival and reducing infection rates in pandemic regions. Here, we report an ultrasensitive and instrument-free EBOV detection assay based on colloidal carbon immunochromatography. Carbon nanoparticle-labeled rabbit anti-EBOV-VP40 IgG were concentrated in the conjugate pad, monoclonal antibody (McAb, 4B7F9) against EBOV-VP40 and goat anti-rabbit IgG were immobilized on the nitrocellulose membrane with 2 μL/cm at a concentration of 1 mg/mL as test and control lines, respectively. Then the sample application pad, conjugate release pad, nitrocellulose membrane and absorbent pad were assembled into a lateral flow test strip. The test strip shows strong specificity against related viruses that share similar clinical symptoms and geographic range with EBOV, including marburg virus, influenza virus, yellow fever virus and dengue virus. In addition, 1 500 negative serums were tested with false-positive rate of 1.3‰ which significantly lower than that of ReEBOV™ colloidal gold test kit recommended by World Health Organization (WHO). The sensitivity of this strip was analyzed using inactivated EBOV with detection limit of 100 ng/mL (10⁶ copies/mL) which clearly higher than that of ReEBOV™ dipstick (10⁸ copies/mL). Furthermore, the strip showed excellent thermal stability characteristics in room temperature and could be as a point-of-care (POC), ultra-sensitive and specific promising candidate for EBOV serological screening in rural Africa or entry/exit ports.
Animals ; Carbon ; Ebolavirus ; Hemorrhagic Fever, Ebola ; Humans ; Nanoparticles ; Rabbits

Since the first case of Ebola virus disease (EVD) in Guinea was reported in March 2014 by World Health Organization (WHO), the outbreak has continued through the year and the total number of 19,065 patients was reported as the confirmed or suspected in the EVD-affected countries. Among the cases, 7,388 patients were reported death by 19 December. Currently, available therapeutics to treat the infected patients or vaccines to prevent people from infection is not developed yet while viral diagnostic methods were already developed and firmly established in a lot of countries as a first step for the preparedness of Ebola outbreak. Some potential therapeutic materials including ZMapp were supplied and the treated people got over the EVD. Several candidates of vaccines also were investigated their efficacy in animal models by National Institute of Health (NIH) and Department of Defense, and they are processing of clinical tests in West Africa aiming to finish the development by the 2015. Vaccine and therapeutic development is essential to stop the EVD outbreak in West Africa, also to protect the world from the risk which can be generated by potential spread of Ebola virus.
Africa, Western ; Ebolavirus ; Guinea ; Hemorrhagic Fever, Ebola ; Humans ; Models, Animal ; Vaccines ; World Health Organization

Africa ; Asian Continental Ancestry Group ; Hemorrhagic Fever, Ebola ; Humans ; Military Personnel

China ; Hemorrhagic Fever, Ebola ; prevention & control ; Humans ; Influenza, Human ; epidemiology ; prevention & control ; Research