Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South FloridaMarine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida

RESEARCH - SENSOR RESEARCH
NASBA ASSAY FOR ENTEROVIRUSES


Poliovirus is just one example of the many enteroviruses present in wastewater and the environment.
Enteroviruses are a group of small RNA viruses in the family Picornaviridae that are responsible for a wide range of symptomatic and asymptomatic infections worldwide. Enteroviruses are second only to rhinoviruses as the most common infectious viral agent in humans in the US (http://www.cdc.gov/ncidod/dvrd/revb/
enterovirus/non-polio_entero.htm
). The Centers for Disease Control and Prevention (CDC) estimate that there are 10-15 million symptomatic infections in humans in the United States each year, which includes 90% of all viral meningitis cases (http://www.cdc.gov/ncidod/dvrd/revb/
enterovirus/non-polio_entero.htm
). The majority of infections, however, are asymptomatic or result in only a mild cold or flu-like illness that may be significantly underreported. In the rare instances involving more serious infections, illnesses such as myocarditis, encephalitis and paralysis, as well as meningitis, can occur. In addition, enteroviral infections have been linked to the occurrence of type I diabetes and therefore may play a larger role in overall health of exposed populations (Gamble 1969; Clements et al., 1995; Andreoletti et al., 1997).

Rapid and sensitive detection of enteroviruses is important in both clinical samples to diagnose illness and in environmental samples to assess risk of wastewater contamination and potential health hazards. We have developed a rapid, sensitive, and specific assay for the detection and quantification of enteroviruses using nucleic acid sequence-based amplification (NASBA). The inclusion of an internal control (IC) increased the precision and accuracy of the method over a standard NASBA assay as well as provided a way to detect assay inhibition. The assay was sensitive to 10 viral particles with amplification and detection occurring in as little as 18 minutes. The assay detected a variety of different enteroviruses to the exclusion of non-target viruses.


Table 1. Specificity of the enteroviral NASBA assay toward target and non-target viruses.



Typical amplification plots for viral concentrations from 104 to 10 particles (a-d, respectively). Thick black line represents the wild type (PV) amplification. Grey line represents the internal control amplification. Horizontal line is the detection threshold, set at 1.3.


References:

Andreoletti, L., Hober, D., Hober-Vandenberghe, C., Belaich, S., Vantyghem, M.C., Lefebvre, J. and Wattré, P., 1997. Detection of Coxsackie B virus RNA sequences in whole blood samples from adult patients at the onset of type I diabetes. J. Med. Virol. 52, 121-127.

Casper, E.T., S. S. Patterson, M.C. Smith, and J.H. Paul. 2004. Development and Evaluation of a Method to Detect and Quantify Enteroviruses Using NASBA and Internal Control RNA (IC-NASBA). J. Virol. Meth. (in press).

Clements, G.B., Galbraith, D.N. and Taylor, K.W., 1995. Coxsackie B virus infection and the onset of childhood diabetes. Lancet 346, 221-223.

Gamble, D.R. and Taylor, K.W., 1969. Viral antibodies in diabetes mellitus. Br. Med. J. 3, 631-633



Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Copyright © 2008, University of South Florida, College of Marine Science 140 7th Avenue S., St. Petersburg, FL 33701 Marine Microbiology Group - College of Marine Science - University of South Florida
Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida Marine Microbiology Group - College of Marine Science - University of South Florida