title: The development of a multiplex real-time RT-PCR for the detection of adenovirus, astrovirus, rotavirus and sapovirus from stool samples.
abstract: Viral gastroenteritis is a major health problem with significant morbidity and economic consequences. Viral gastroenteritis is caused by a number of viruses, including norovirus, rotavirus, adenovirus, astrovirus, and sapovirus. Conventional diagnosis is based on direct antigen detection and electron microscopy, however enzyme immunoassay's are insensitive and not available for all relevant pathogens, and electron microscope (EM) is no longer routinely carried out in most laboratories. Most laboratories now offer norovirus real-time PCR testing however the availability of other assays is variable. Commercial methods for the detection of inflectional intestinal disease (IID) are available but these can be expensive and are not commonly used. This paper describes the development of a single multiplex assay for the simultaneous detection of adenovirus, astrovirus, rotavirus and sapovirus from stool samples. The multiplex was evaluated by assessing endpoint sensitivity, specificity, panel of clinical samples, quality control (QC) panel and the robustness and reproducibility of the multiplex.
authors.affiliation: West of Scotland Specialist Virology Centre, United Kingdom. Electronic address: susan.bennett@ggc.scot.nhs.uk.
authors.forename: Susan
authors.initials: S
authors.lastname: Bennett
authors.affiliation: West of Scotland Specialist Virology Centre, United Kingdom.
authors.forename: Rory N
authors.initials: RN
authors.lastname: Gunson
mesh_terms: D000258:Adenovirus Infections, Human / Q000175:diagnosis / Q000821:virology; D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification*; D002139:Caliciviridae / Q000235:genetics / Q000302:isolation & purification*; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000821:virology; D017931:DNA Primers; D015342:DNA Probes; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007124:Immunoenzyme Techniques; D057230:Limit of Detection; D001257:Mamastrovirus / Q000235:genetics / Q000302:isolation & purification*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis / Q000821:virology; D029341:Sapovirus / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D015342:DNA Probes
keywords: Adenovirus; Astrovirus; Multiplex PCR; Rotavirus; Sapovirus; Viral gastroenteritis