- Ultrasensitive qualitative detection of chikungunya virus by
reverse transcription coupled real time polymerase chain
Chikungunya virus is transmitted by mosquito. The most common
symptoms of chikungunya infection are fever and joint pain,
which can be excruciating, leading to the colloquial name
“breakbone fever.” Other symptoms may include headache, muscle
pain, joint swelling, and rash. Chikungunya means "that which
bends up" or "to be contorted" in the Kimakonde language and was
first described in the 1950s in Tanzania. Chikungunya is an RNA
virus in the Alphavirus genus of the Togaviridae family.
In 2013, chikungunya viral infection was reported in humans in
the Caribbean. The virus quickly spread to almost every island,
with many cases found in the Dominican Republic and Haiti. In
2014, cases of this viral infection started to be reported in
Central and South America and it quickly became endemic. As of
September 5, 2014, the Pan American Health Organization had
confirmed 8210 cases and 37 deaths resulting from chikungunya
viral infection, with 751 reported cases in the continental
United States, mostly in southern Florida.
Viral infection of nonhuman primates is possible; serological
survey of wild-caught monkeys in Southeast Asia has shown
widespread exposure to this virus. The first report of
chikungunya virus isolation from non-human primates was
published in 2009 in Malaysia (Apandi et al., 2009), and
cross-transmission between human and monkey through mosquitoes
was then proposed.
Diagnosis of chikungunya viral
infection is mostly based on serological and PCR techniques.
Viral culture followed by viral antigen detection is a sensitive
method but must be performed under BSL3 biosafety conditions,
and is time consuming and labor intensive, limiting its
usefulness. Serological methods are reliable but are not
appropriate in early stage infection, i.e., before 5–6 days
after clinical onset, because antibody titer is low during early
stages of infection and serology is not sensitive enough to
detect small changes in antibody titer. Nucleic acid
amplification by PCR is an appropriate diagnostic tool at an
early stage of infection, while the patient is viremic (Laurent
et al., 2007).
Help confirm the disease causing agent
Help ensure that animal colonies are free of this virus
Early prevention of spread of this virus among a colony
Minimize personnel exposure to this virus
Safety monitoring of biological products and vaccines that
derive from primates
Apandi, Y., Nazni, W.A., Noor Azleen, Z.A.,
Vythilingam, I., Noorazian, M.Y. , Azahari, A.H., Zainah, S. and
Lee, H.L. (2009) The first isolation of chikungunya virus from
nonhuman primates in Malaysia. J. Gen. Mol. Virol. 1: 035–039
Laurent, P., Le Roux, K., Grivard, P., Bertil, G., Naze, F.,
Picard, M., Staikowsky, F., Barau, G., Schuffenecker, I. and
Michault, A. (2007) Development of a sensitive real-time reverse
transcriptase PCR assay with an internal control to detect and
quantify Chikungunya virus. Clin. Chem. 53: 1408–1414.
0.2 ml whole blood in EDTA (purple top) or ACD (yellow top) tube,
or 0.2 ml plasma or serum, or 0.2 ml cerebrospinal fluid, or 0.2
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Qualitative reverse transcription coupled real time PCR