ABSTRACT
BACKGROUND: Recent outbreaks of poliomyelitis caused by vaccine-derived virus have raised concerns that vaccine-derived poliovirus may continue to circulate after eradication. In these outbreaks, the virus appears to have replicated for > or =2 years before detection. Early detection is critical for an effective response to these outbreaks. Although acute flaccid paralysis (AFP) surveillance will remain the standard for poliovirus detection, wastewater sampling could be a useful supplement. In this study, we evaluated the sensitivity of wastewater sampling by concurrently collecting stools from children aged < 3 years attending two neighbourhood clinics in Havana, Cuba, and wastewater from the same neighbourhoods. METHODS: Sample collection was begun during the third week after the national immunization campaign, continued weekly through the seventh week, and was repeated during weeks 15 and 19. Virus detection and titration were performed using both cell culture and polymerase chain reaction techniques. RESULTS: Wastewater sampling was found to be at least as sensitive as stool sampling under these conditions. Poliovirus was isolated from children through week 7, suggesting that viral shedding reached undetectable levels between weeks 8 and 14. The last virus-positive wastewater sample was collected during week 15. CONCLUSIONS: Wastewater sampling under the conditions studied can be a sensitive supplement to AFP surveillance. Similar studies under different conditions are needed to determine the role of wastewater sampling in post-eradication surveillance.
Subject(s)
Environmental Monitoring/methods , Feces/virology , Mass Vaccination , Poliovirus/isolation & purification , Sewage/virology , Child, Preschool , Cuba , Humans , Infant , Infant, Newborn , Poliomyelitis/prevention & control , Poliovirus/classification , Poliovirus Vaccines/adverse effects , Polymerase Chain Reaction/methods , Population Surveillance , Specimen Handling/methodsABSTRACT
For the detection of Enterovirus, we devised a direct economical method of polymerase chain reaction which does not require a previous extraction of ribonucleic acid and uses infected cell culture supernatants. The system was developed on the basis of universal primers of Enterovirus and specific primers of vaccinal strain Sabin 1. The achieved results proved that the non-existence of methods of RNA extraction and purification prior to the reaction does not affect the susceptibility and specificity of the system, in the rapid detection of Enterovirus genomes and identification of vaccinal strains of poliovirus.
Subject(s)
Humans , Enterovirus , Polymerase Chain Reaction/methods , Base Sequence , DNA, Viral , Enterovirus , Enterovirus Infections/diagnosis , Enterovirus Infections/virology , Meningoencephalitis , Molecular Sequence Data , RNA, Viral , Sensitivity and SpecificityABSTRACT
This paper reported the appraisal of a novel technology for the detection of enterovirus in sewage based on a viral recovery method using polyethylenglycol as a concentrating agent and on the combined use of viral isolation and polymerase chain reaction as viral detection and identification techniques. It was also confirmed that the viral recovery method is highly efficient since it allows to recover all viruses present in sewage and to preserve their infective capacity. It was proved that the polymerase chain reaction (PCR) is less sensitive than cell culture for detecting enteroviruses in sewage. It is also possible to use guadinine isothiocyanate, whose components can be prepared in the lab, as an alternative method for taking out and purifying nucleic acids instead of using the conventional TRIZOL method which is the one recommended in these cases by the World Health Organization.
