A simplified cytomegalovirus pp65 antigenemia assay procedure.

A simplified cytomegalovirus (CMV) pp65 antigenemia assay using a one-step erythrocyte lysis, fixation and permeabilization process was compared with a standard protocol, the CMV CINAkit (Argene Biosoft) assay. The results were comparable, both quantitatively and qualitatively. The new method saves time. It also provides flexibility because the cell suspension can be stored so that test completion can be deferred if so desired.

Are waterborne astrovirus implicated in acute digestive morbidity (E.MI.R.A. study)?

BACKGROUND: With rotavirus and Norwalk-like viruses, astroviruses are now recognized as important etiologic agents of viral gastroenteritis in all age groups. However, astrovirus is neither routinely screened for in stool samples, nor in environmental samples, and data on the health impact of waterborne astrovirus are lacking. OBJECTIVES: To assess the potential impact of astrovirus in drinking water on the incidence of acute digestive conditions (ADC) among a panel of volunteers. STUDY DESIGN: The Epidemiology and MIcrobial Risk Assessment (E.MI.R.A.) study combined a daily epidemiological follow-up of digestive morbidity among a panel of 544 volunteers supplied by French public water systems, and a microbiological surveillance of drinking water. Cases of digestive morbidity were collected through weekly telephone calls. The bacterial, virological and parasitic quality of tap water was assessed monthly. Additional samples were collected if the incidence of ADC increased. The relationship between incidence of ADC during a 7-day period centered about the water sampling day and astrovirus RNA prevalence in drinking water was modeled by regression techniques, taking into account several confounders. RESULTS: 12% (8/68) of the analyzed water samples were positive for astrovirus, and presence of astrovirus RNA was associated with a significant increased risk of ADC: RR=1.51 (95% CI=[1.17-1.94], P value=0.002). CONCLUSIONS: This result suggests a role for waterborne astrovirus in the endemic level of digestive morbidity in the general population. Perhaps astrovirus is a candidate test target for viral surveillance of drinking water.

A new analytical tool to assess health risks associated with the virological quality of drinking water (EMIRA study).

This work assessed the risks associated with the virological quality of tapwater using a molecular analytical tool manageable in a field survey. It combined a daily epidemiological follow-up of digestive morbidity among a panel of volunteers and a microbiological surveillance of drinking water. RT-PCR was used for detection of enterovirus, rotavirus and astrovirus. 712 cases of acute digestive conditions occurred in the 544 volunteers. 38% (9/24) raw water and 23% (10/44) tap water samples were positive for at least one virus marker with 9/10 positive tap water samples complying with bacterial criteria. No statistically significant association was found between the presence of viral markers and observed incidence of digestive morbidity. However, when an outbreak occurred, enterovirus and rotavirus RNA was detected in the corresponding stored tap water samples. Sequencing of the amplified fragments showed that the rotavirus detected was of bovine origin. This work demonstrated that enteric virus markers were common in tapwater of the study communities (characterised by a vulnerable raw water) despite absence of bacterial indicators. Tangential ultrafiltration coupled to RT-PCR allowed a simultaneous and fast detection of the study viruses from environmental samples. This process is a promising tool usable for virological water surveillance, in as much the corresponding know-how is transferred to the field professionals.

Detection of human and animal rotavirus sequences in drinking water.

Reverse transcription-PCR analysis of drinking water in the homes of 56 children suffering from rotaviral gastroenteritis has shown the presence of the rotavirus genome in four samples. These strains were different from human rotaviruses detected in the children's feces, as determined by sequencing of the VP7-amplified fragments-three of them of animal origin (porcine or bovine) and one of human origin.

Monitoring rotavirus environmental contamination in a pediatric unit using polymerase chain reaction.

Rotavirus environmental contamination in a pediatric unit was investigated. Surfaces were swabbed, then viruses eluted, ultracentrifuged, and detected by polymerase chain reaction (PCR) amplification. Of 55 samples, 25 (46%) tested positive. Rotavirus RNA was more prevalent on surfaces in direct contact with children (thermometers and play mats) than on other environmental surfaces (washbasins, door handles, etc). PCR has proved useful for monitoring rotavirus environmental contamination.

[Virus resistance in a hospital environment: overview of the virucide activity of disinfectants used in liquid form]. / Résistance des virus dans l'environnement hospitalier: le point sur l'activité virucide des désinfectants utilisés à l'état liquide.

Human pathogenic viruses can be detected in the hospital environment, on contaminated surfaces or medical instruments. Their transmission to patients or staff has already been reported. Lipophilic viruses (HIV, HBV, HCV,...) are susceptible to many liquid chemicals, but they can survive during short time on inadequately disinfected surfaces. Hydrophilic viruses, without envelope, are more resistant, but generally not associated with severe illnesses. Viruses survival in environment depends on many factors and is always improved with viral aggregation and low temperature, whereas organic matters and relative humidity effects are contrasted. The mechanism of virucide disinfectants is not yet well established, and their targets are not known with precision. Different disinfection procedures (disinfectant concentration, contact time, temperature, pH) can provide a similar virucidal activity on a given virus. The virucidal activity of a disinfectant is evaluated with a cell culture assay in Afnor guidelines. But, there are three major problems with this method, concerning need of high viruses titers, residual disinfectant cytotoxicity on cell culture, and non cultivable viruses. Non standardized tests are also described in papers, but their results can generally not be compared. Molecular biology improvements may lead to reproducible and sensitive tests. At present, no general disinfection procedure effective for most of the viruses, without risks for staff or materials, and with an acceptable economic cost can be recommended.

[Cytomegalovirus infection in pregnant women. Seroepidemiological prospective study in 1,018 women in Isere]. / Infection à cytomégalovirus chez la femme enceinte. Etude séro(-)épidémiologique prospective chez 1,018 femmes en Isère.

Cytomegalovirus (CMV) is the leading cause of viral congenital infections. In children, the consequences may be severe, especially in case of maternal primary infection during pregnancy. A prospective study was carried out in the department of Isère, in 1,018 pregnant women, in order to establish the seroprevalence of CMV, the frequency of primary infections during pregnancy and the associated risk factors. The overall seroprevalence was 51.5%; it increased significantly with age, parity, and low socioeconomic status. It was higher in women born in the South of France (51.6%) than in those born in the North (37.4%). Among a total of 878 women with serological follow-up, 7 primary infection cases (0.8%) were observed. Seventeen women (1.9%) presented border IgM values in the first serum, and these values were not related to recent infection. Extrapolation of the results to the whole department of Isère, suggests that each year about 100 pregnant women would be concerned by CMV primary infection, with 2 or 3 cases of death or severe sequelae in children. In light of these results, the interest of serological screening is discussed.

Cytomegalovirus seroprevalence in French pregnant women: parity and place of birth as major predictive factors.

In this study we present data on cytomegalovirus (CMV) seroprevalence in pregnant women in France. One thousand and eighteen women were enrolled in a prospective study carried out in Grenoble. The overall rate of seropositivity, using a specific IgG ELISA test, was 51.5 % (95 % CI: 48.5-54.5). Among a homogeneous population of 873 women born in France with high or middle socioeconomic status, CMV seropositivity increased with age and parity. The seroprevalence according to age was found to depend on parity. It increased with age in women with no children or with only one; it was higher but no more age-dependent in women with two children or more. In addition, CMV seroprevalence was significantly higher in women born in southern France (51.6%) than in those born in northern France (37.4%), these findings being consistent with the existence, within France, of a gradient in seroprevalence rate, increasing from the North to the South. A logistic regression analysis reveals the place of birth in France as a major predictive factor of CMV antibody status (OR: 3.5) followed by age (OR: 2) and parity (OR: 1.7). In this study, we show an independent effect of parity on CMV seroprevalence, arguing for the importance of child-to-mother transmission; nevertheless, the latitude of the place of birth, even within a size-limited country such as France, appears to be a major predictive factor of CMV seroprevalence.

Multicenter evaluating of a commercially available PCR assay for diagnosing enterovirus infection in a panel of cerebrospinal fluid specimens.

Thirteen laboratories participated in blind tests of a panel of 20 coded cerebrospinal fluid specimens (7 uninfected samples, 3 samples infected with 1 50% tissue culture infective dose [TCID50]/0.1 ml [nonenterovirus strains], and 10 samples infected with 10, 1, or 0.1 TCID50/0.1 ml [three different enterovirus serotypes]) on the Amplicor enterovirus PCR assay (Roche Diagnostic Systems). The panel was also evaluated by in-house PCR (two nested-PCR and three one-step PCR assay) or tissue culture (eight laboratories). The viral load was shown to influence greatly the sensitivity of the assay. The average sensitivity of the Amplicor test ranged from 67 to 98% for viral titers of 1 to 10 TCID50/0.1 ml, respectively; titers of 0.1 TCID50/0.1 ml resulted in a sensitivity of only 16%. The overall specificity of the Amplicor test was 98%. The Amplicor assay compared favorably to the five in-house PCR tests (no significant difference in either sensitivity or specificity) and was much more sensitive than tissue culture (P < 0.001), even for high viral loads. It was easy to perform, rapid (about 6 h), well-standardized, and appeared to be suitable for the diagnosis of enterovirus meningitis on a routine basis in laboratories trained in molecular biology techniques.