Ultrasensitive qualitative detection of Ebola virus
(including Reston, Mayibout, Mavinga, Zaire, Gabon, Sudan, and Cote
d'Ivoire strains) by reverse
transcription coupled real time polymerase chain reaction
hemorrhagic fever (VHF) is a clinical syndrome caused by a
number of different viruses. These viruses include members of
the Filoviridae family (Marburg virus [MBGV] and Ebola virus
[EBOV]), Arenaviridae family (Lassa virus [LASV] and Junin,
Machupo, Sabia, and Guanarito viruses), Bunyaviridae family
(Crimean-Congo hemorrhagic fever virus [CCHFV], Rift Valley
fever virus [RVFV], and Hanta viruses), and Flaviviridae family
(yellow fever virus [YFV] and dengue virus [DENV]). The natural
reservoirs of these viruses are arthropods, ticks, and rodents
but the reservoir of filoviruses is not known.
by these hemorrhagic viruses can result in a wide spectrum of
clinical manifestations such as diarrhea, myalgia, cough,
headache, pneumonia, encephalopathy, and hepatitis. Hemorrhage
is the characteristic manifestation, although nonhemorrhagic
infections are also common. The mortality rate of VHF infection
is very high. Filoviruses, arenaviruses, and CCHFV are of
particular relevance because they can be transmitted from human
to human, thus causing epidemics with high mortality rates.
VHF viruses, Ebola virus (EBOV) is of great concern as several
outbreaks of this viral infection have been reported (Volchkov
et al., 1997). This virus can cause severe hemorrhagic fevers in
humans and nonhuman primates. Outbreaks such as the latest ones
in the Democratic Republic of the Congo (MBGV) and Uganda (EBOV)
are unpredictable and pose considerable public health concern to
the affected as well as neighboring countries. In April 1996,
laboratory testing of imported nonhuman primates (as mandated by
quarantine regulations) identified 2 cynomolgus macaques (Macaca
fascicularis) infected with Ebola (subtype Reston) virus in a
US-registered quarantine facility (Rollin et al., 1999). The
animals were part of a shipment of 100 nonhuman primates
imported from the Philippines. Two additional infected
animals, thought to be in the incubation phase, were also
identified among the remaining 48 animals in the affected
Recent studies have found that Ebola Reston
can infect pigs, and there is concern that pigs could serve as
an interim or amplifying host for ebolaviruses (Haddock et al.,
syndrome of EBOV hemorrhagic fever is characterized by
generalized fluid distribution problems, hypotension,
coagulation disorders, variable degrees of hemorrhage, and
widespread focal tissue destruction. Morphological studies on
postmortem material indicate that mononuclear phagocytic cells
are the primary targets for filovirus replication. There are two
species of EBOV: the Zaire species shows the highest mortality
in humans, whereas the Reston species may be apathogenic, based
on very limited data.
absence of bleeding or organ manifestation, VHF is clinically
difficult to diagnose, and the various etiologic agents of VHF
can hardly be differentiated by clinical tests. The definitive
diagnosis of a VHF relies mainly on laboratory testing and it is
important to identify the causative viral agent to initiate the
appropriate treatment and infection control procedures.
Serological testing of EBOVs is not practical because patients
usually die before developing antibodies, necessitating rapid
virus detection. Detection of EBOVs using a molecular biology
approach provides the best alternative for rapid diagnosis of
Help confirm the disease causing agent
Help ensure that animal colonies are free of Ebola virus
Early prevention of spread of the virus among a colony
Minimize personnel exposure to the virus
Safety monitoring of biological products and vaccines
that derive from primates
Leroy EM, Baize S, Lu CY, McCormick JB, Georges AJ, Georges-Courbot
MC, Lansoud-Soukate J, Fisher-Hoch SP. (2000) Diagnosis of Ebola
haemorrhagic fever by RT-PCR in an epidemic setting. J. Med.
Virol. 2000 60:463-7.
Rollin, P.E., Williams, R.J., Bressler,
D.S., Pearson, S., Cottingham, M., Pucak, G., Sanchez, A.,
Trappier, S.G., Peters, R.L., Greer, P.W., Zaki, S., Demarcus,
T., Hendricks, K., Kelley, M., Simpson, D., Geisbert, T.W.,
Jahrling, P.B., Peters, C.J. and Ksiazek, T.G.(1999) Ebola
(subtype Reston) virus among quarantined nonhuman primates
recently imported from the Philippines to the United States. J.
Infect. Dis.. 179 Suppl 1:S108-14.
Volchkov, V., Volchkova,
V., Eckel, C., Klenk, H.D., Bouloy, M., LeGuenno, B. and
Feldmann, H. (1997) Emergence of subtype Zaire Ebola virus in
Gabon. Virology 232:139-44
Haddock, E. et al. (2020)
Reston virus causes severe respiratory disease in young domestic
pigs. PNAS DOI: 10.1073/pnas.2015657118.
Specimen requirements: 0.2 ml whole blood in EDTA (purple top) tube, or 0.2 ml plasma or serum.
Contact Zoologix if advice is needed to determine an appropriate specimen type for a specific diagnostic application. For specimen types not listed here, please contact Zoologix to confirm specimen acceptability and shipping instructions.
specimen types, if there will be a delay in shipping, or during
very warm weather, refrigerate specimens until shipped and ship
with a cold pack unless more stringent shipping requirements are
specified. Frozen specimens should be shipped so as to remain
frozen in transit. See shipping
instructions for more information.
2 business days
Qualitative reverse transcription coupled real time PCR