primate assay data sheet
Enterovirus
Test code:
S0058 -
Ultrasensitive qualitative detection of enterovirus by reverse
transcription coupled real time polymerase chain reaction
The
enteroviruses (EVs) are among the most common and most important
viral pathogens of humans, and EVs also infect nonhuman
primates. The EVs comprise 67 distinct serotypes within the
family Picornaviridae. Most EV infections are asymptomatic. For
symptomatic patients, the most common manifestation of EV
infection may only be a nonspecific febrile illness with or
without a rash. This so-called viral syndrome is one of the most
common causes of fever among human children. Infected patients
often develop upper respiratory symptoms very similar to illness
caused by rhinoviruses in the winter months, making diagnosis
very difficult.
Enteroviruses are the most common cause of meningitis in humans
in the United States. It is also likely that EV infection in
primates is a major cause of aseptic meningitis. Diagnosis of EV
meningitis can be difficult, especially in young infant
primates, because meningitis due to bacterial and other viral
infection can present very similar symptoms. Even though EV
meningitis is often benign in outcome and no specific therapy is
indicated, EV-infected animals may be given unnecessary
antibiotic and antiviral therapies due to misdiagnosis.
Laboratory
diagnosis of enteroviruses is traditionally performed by viral
culture and recognition of cytopathic effects, requiring a high
degree of expertise and labor. Some EV serotypes, particularly
within the coxsackievirus A group, do not grow at all in cell
culture. Of greater concern is that 25 to 35% of specimens from
human patients with characteristic EV infections are negative by
cell culture (Chonmaitree et al., 1988) because of antibody
neutralization in situ. Even though some EVs may grow in cell
culture, they often grow very slowly, sometimes taking a week or
more before any cytopathic effect can be observed.
Serological
testing of EVs is not practical because there is no commonly
shared antigen among the different EV serotypes (Herrmann et
al., 1979). Detection of EVs using a molecular approach provides
the best opportunity for rapid diagnosis of EVs in primates,
because consensus sequences among different serotypes of EVs are
available in the untranslated regions of EVs. Enterovirus
detection by PCR over these consensus sequences thus provides
the most rapid, sensitive and specific method for the diagnosis
of this infection. The technique also helps eliminate false
negative diagnoses.
Utilities:
-
Help confirm the disease causing agent
-
Help ensure that animal colonies are free of Enteroviruses
-
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
References:
Chonmaitree, T., Ford, C., Sanders, C. and Lucia, H.L. (1988)
Comparison of cell culture for rapid isolation of enteroviruses.
J. Clin. Microbiol. 26:2576-2580.
Herrmann, J.E., Hendry,
R.M. and Collins, F. (1979) Factors involved in enzyme-linked
immunoassay of viruses and evaluation of the method for
identification of enteroviruses. J. Clin. Microbiol. 10:
210-217.
Specimen requirements: 0.2 ml whole blood in EDTA (purple top) tube, or 0.2 ml CSF, urine, plasma, serum or nasal wash, or
rectal swab, or throat swab, or 0.2 ml fresh or frozen tissue.
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.
For all
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.
Turnaround time:
2 business days
Methodology:
Qualitative reverse transcription coupled real time PCR
Normal range:
Nondetected