[Video: An innovation for developmental care in the neonatal intensive care unit]. / La surveillance vidéo : une innovation au service des soins de développement en unité de soins intensifs de néonatologie.

INTRODUCTION: Our neonatal intensive care unit (NICU) was designed with a web-camera system in eight private rooms and four open wards. This equipment was installed to both guarantee children's safety in the NICU and promote the bond between the child and his/her family through a viewing service permitted by an Internet access. METHOD: We evaluated the web-camera system in its 5th year with two types of users. The nursing staff was asked about use of the video and its impact on their management of NICU patients. Questionnaires for parents sought to determine how they used the system and their feelings about it. RESULTS: Ninety-three percent of the nursing staff used the web-camera system to provide medical supervision or to comply with developmental care, mainly to respect the baby's natural rhythm by initiating care only when the baby showed signs of awakening. The web-camera system allowed them to observe the baby, verify his/her good body position, and spot discomfort situations. It helped provide a faster and personalized answer that was adjusted to the baby's needs. Sixty-one percent of the parents used the remote connection, half of them to present the child to the family. Only 17% of parents were embarrassed by the cameras; 88% of parents thought that the video was an additional safety device in NICU management. CONCLUSION: The web-camera system appeared to be an interesting technology in the NICU to support developmental care when children could be put in private rooms. It helped staff react faster to situations that created an inappropriate stimulation for the baby. It also allowed to them to respect the baby's natural rhythm. It reassured nursing staff and parents and it facilitated the baby's integration into the family.

Shift of the 2009-2011 measles outbreak from children to adults: an observational review at the University Hospital of Clermont-Ferrand, France.

In 2009-2011, 113 adult in- and outpatients with measles were referred to the University Hospital of Clermont-Ferrand (centre of France): 71 (62.8 %) needed hospitalisation, 31 had pneumonia, 29 diarrhoea, 47 liver enzymes elevation, 38 thrombopaenia, one encephalitis and there were no deaths. Nineteen cases occurred among healthcare workers and five of them were hospital-acquired. There were 92 unvaccinated patients. The 2011 peak of that measles re-emerging epidemic occurred when non-immunised adults were affected.

Repeated genomic transfers from echovirus 30 to echovirus 6 lineages indicate co-divergence between co-circulating populations of the two human enterovirus serotypes.

Human echovirus types 6 (E-6) and 30 (E-30) cause seasonal epidemics of aseptic meningitis. These two enteroviruses are frequently observed in co-circulation, an epidemiological pattern that is prerequisite for the occurrence of dual infections, which can lead to recombination between co-infecting virus strains. Viral sequences were determined at loci 1D (VP1 capsid protein) and 3CD (non structural proteins) in 49 E-6 strains recovered in a single geographical region in France from 1999 to 2007, during the epidemiological survey of enterovirus infections. They were compared with previously recorded sequences of E-30 strains to investigate their evolutionary histories and possible recombination patterns. Phylogenetic analyses identified two distinct E-6 populations and different subpopulations. Assuming a relaxed molecular clock model and a Bayesian skyline demographic model in coalescent analyses with the BEAST program, the substitution rate in E-6 was estimated at 8.597×10(-3) and 6.252×10(-3) substitution/site/year for loci 1D and 3CD respectively. Consistent estimates of divergence times (t(MRCA)) were obtained for loci 1D and 3CD indicating that two distinct E-6 populations originated in 1997 and 1999. Incongruent phylogenetic patterns inferred for the two loci were indicative of recombination events between the two populations. Phylogenies including the E-30 3CD sequences showed close genetic relationships between E-6 and discrete E-30 subpopulations. Recombination breakpoints were located with statistical significance in E-6 and E-30 genomes. Estimates of t(MRCA) of phylogenetic recombinant clades indicated directional genetic transfers from E-30 to E-6 populations and their co-divergence over the time period studied.

Heat shock cognate 70 protein secretion as a new growth arrest signal for cancer cells.

Earlier studies indicated that density-arrested cancer cells released an unidentified growth inhibitor whose secretion was prevented by overexpression of the lysosomal protease cathepsin D (cath D). In this study, this growth inhibitor was purified by affinity chromatography and identified as the heat shock cognate 70 protein (hsc70) based on its peptide microsequencing and specific antibody recognition. Among intracellular proteins, including other heat shock proteins, only constitutive hsc70 was secreted in response to the high-cell density. Moreover, hsc70 secretion from cancer cells was generated by serum deprivation, whereas its cellular concentration did not change. Prevention of Hsc70 secretion by cath D overexpression was associated with the formation of multilayer cell cultures, thus indicating a loss of contact inhibition. In addition, we showed that supplementing the culture medium with purified hsc70 inhibited cell proliferation in the nanomolar range. Conversely, removal of this extracellular hsc70 from the medium by either retention on ADP-agarose or competition at the Hsc70 binding site restored cell proliferation. Hsc70 appears active in human breast cancer cells and hypersecreted by direct cath D inhibition. These results suggest a new role of this secreted hsc70 chaperone in cell proliferation that might account for the higher tumor growth of cancer cells overexpressing cath D.

Impact of rapid enterovirus molecular diagnosis on the management of infants, children, and adults with aseptic meningitis.

Enteroviruses (EV) are the main etiological agents of aseptic meningitis. Diagnosis is made by detecting the genome using RT-PCR. The aim of the study was to evaluate the impact of a positive diagnosis on the management of infants, children, and adults. During 2005, 442 patients were admitted to hospital with suspected meningitis. Clinical and laboratory data and initial treatment were recorded for all patients with enteroviral meningitis. The turnaround time of tests and the length of hospital stay were analyzed. The results showed that EV-PCR detected EV in 69 patients (16%), 23% (16/69) were adults. About 18% of CSF samples had no pleocytosis. After positive PCR results, 63% of children were discharged immediately (mean 2 hr 30 min) and 95% within 24 hr. Infants and adults were discharged later (after 1.8 and 2 days, respectively). The use of antibiotics was significantly lower in children than in infants and adults. The PCR results allowed discontinuation of antibiotics in 50-60% of all patients treated. Patients received acyclovir in 16% of cases (7% children vs. 50% adults) and 23% (11% vs. 69%) underwent a CT scan. Clinical data were compared between patients whose positive EV-PCR results were available within 24 hr (n = 32) and those whose results were available > 24 hr after collection of CSF (n = 14). Duration of antibiotic treatment (difference: 2.3 days; P = 0.05) was reduced between the two groups. No statistical difference in the length of stay was observed. The EV-PCR assay should be performed daily in hospital laboratory practice and considered as part of the initial management of meningitis.

Phylogeography of circulating populations of human echovirus 30 over 50 years: nucleotide polymorphism and signature of purifying selection in the VP1 capsid protein gene.

A comprehensive set of 443 1D gene sequences (encoding the VP1 capsid protein) was analyzed to investigate the phylogenetic relationships and evolutionary patterns among strains of human echovirus 30 (E30; genus Enterovirus, family Picornaviridae) characterized over 50 years. Maximum-likelihood (ML) phylogenetic trees of complete and nonredundant 1D gene sequences (total length=876 nucleotides) showed evidence of distinct lineages related to the isolation period of virus strains. Virus transportation was confirmed as a major epidemiological factor in the appearance of epidemics since recurrence of aseptic meningitis outbreaks in a given geographic area was associated with distinct E30 variants detected earlier in distant regions. Detection of the codon changes associated with E30 evolution was investigated with methods implemented in the Datamonkey web server. Evolution of the 1D gene was dominated by continual negative (purifying) selection against nonsynonymous substitutions at most codon sites, as determined by dN/dS ratio. Amino acid polymorphism was maintained at a limited number of sites (10/292) in the VP1 protein (within loops connecting beta strands and C-terminus). Amino acid changes are allowed at these sites because they are likely exposed on the virion particle and nonsynonymous substitutions are observed in the corresponding codons because negative selection is relaxed.

[Genotyping and molecular epidemiology of non polyomyelitic enteroviruses]. / Génotypage et épidémiologie moléculaire des entérovirus non poliomyélitiques.

Nonpolio enteroviruses can be reliably identified with molecular and computer tools for taxonomic, diagnostic and epidemiologic purposes. Seroneutralization tests can efficiently be replaced by genotyping assays using the VP1 capsid protein encoding gene to identify enterovirus strains isolated in cell cultures. Genotyping showed the close genetic relatedness between human enterovirus serotypes and animal enteroviruses and also rhinoviruses currently classified in a separate genus within the Picornaviridae family. Enterovirus genotyping can be done prospectively within 2 to 5 days in a greater number of meningitis patients, using cerebrospinal fluid specimens and hence can help in providing a prompt response to health alert. In the molecular epidemiology of human enteroviruses, recent advances were made by investigating genetic diversity within individual serotypes (genotypes, lineages) and the patterns of circulation and transmission of virus variants involved in epidemics (echovirus 30, enterovirus 71). The observation of epidemiologic features such as the frequent viral immigration of strains from different geographical origins speaks in favour of developing molecular identification of enteroviruses. Recombinant enterovirus strains can also be identified by genotyping. Homologous recombination is a major contributor to the genetic diversity in enteroviruses. Molecular signatures of recombination events are observed in circulating strains, suggesting the occurrence of frequent co-infections during their circulation within the general population. The role of genetic recombination in the emergence of virus variants and its involvement in the epidemiology of human enteroviruses should be investigated.

[From prospective molecular diagnosis of enterovirus meningitis...to the prevention of antibiotic resistance]. / Du diagnostic moléculaire initial prospectif des méningites à entérovirus...à la lutte contre l'antibiorésistance.

Meningitis initially presents with intense manifestations that are not generally specific to a given etiology. The first major question for the physician is to decide whether to initiate a probabilistic treatment. Enteroviruses are a major cause of aseptic meningitis, which is benign in immunocompetent patients. Molecular diagnosis is now becoming the gold standard and its prospective use at the time of patient admission, on the sole basis of clinical suspicion of meningitis, has yielded more reliable data. Cytological and biochemical data from CSF analyses are of low predictive value to influence the initial decision to treat with antibiotics. In addition, cases of meningitis during winter are not uncommon. Adults are concerned in about 25% of cases. Thus, if molecular diagnostic tools are not rapidly available, patient management may be inconsistent, leading to unnecessary scans, laboratory investigations and treatment (including overconsumption of antibiotics). Current progress in the automation and practicability of viral genomic detection yields the result within a few hours after admission. Rapid molecular viral diagnosis of a benign disease that does not require treatment but which is initially worrying is of unquestionable advantage. It is of benefit to both the patient and the community because of its input on health economics, the needless consumption of drugs and, as a result, resistance to antibiotics. The diagnosis of meningitis can no longer remain a retrospective diagnosis after elimination of all the possible causes, since not prescribing unnecessary laboratory tests and not treating are true therapeutic decisions.

[Rapid diagnosis of rotavirus infections: comparative prospective study of two techniques for antigen detection in stool]. / Diagnostic rapide des infections à rotavirus: étude prospective comparative de deux techniques de détection d'antigènes dans les selles.

The ability of two commercially available diagnosis rapid assays in detecting rotavirus antigen was compared in a prospective study conducted from September 2002 to May 2003. Five hundred and twelve faecal specimens were studied by IDEIA Rotavirus enzyme immunoassay test (EIA) and Diarlex MB immunochromatographic test (ICG). Specimens giving discrepant results were examined by electron microscopy (EM) and clinical data reconsidered. Out of 512 stool specimens, 155 (30.3%) were positive and 332 (64.8%) negative with the two assays. Discrepant results were obtained for 25 (4.88%) specimens (24 children, 1 adult), with EIA giving more positive results. The retrospective examination by EM, possible for fifteen stools on the 25 that gave discrepant results, confirmed the presence of rotavirus in 7/14 stools which were positive only by EIA and in the stool specimen that was found positive only by ICG. The 25 clinical observations re-examination showed the presence of GEA signs in all cases. The statistical analysis shows an excellent concordance between the EIA and the ICG tests (kappa = 0.89, IC(95%) = [0.85-0.93]) in spite of the underestimation of ICG test in comparison with EIA test (P < 0.0001).

[Enteroviral meningitis in adults, underestimated illness: description of 30 observations from 1999 to 2000, and evolution of clinical practices during 2001]. / Les méningites à entérovirus chez les adultes. Pathologie sous-estimée ? A propos de 30 observations de 1999 à 2000 et évolution des pratiques médicales en 2001.

Enteroviral meningitis is well documented in children but underestimated in adults. The analysis of 30 cases of adult meningitis prospectively diagnosed by enterovirus genome detection (RT-PCR) in cerebrospinal fluid (CSF) between 1999 and 2000 in routine practice showed diagnosis to be problematic. Characteristic symptoms were inconstant (the association of fever/headache/stiff neck absent in 41%) and sometimes misleading (the presence of peribuccal lesions). CSF data showed a predominance of lymphocytes in only 44% of patients. The most reliable criterion was normal constant CSF glucose levels. Thirty three per cent of patients were admitted during cold months. Management of patients varied markedly between departments, and included computed tomography (33%), and the prescription of aciclovir (20%) or antibiotics (53%). A report of positive enterovirus RT-PCR had only low impact on management because it took 6 days to obtain the results (versus 3 days in children during the same period). These findings were communicated to all hospital physicians concerned and as a result, the number of RT-PCR in adults increased significantly during 2001. Again, enteroviral meningitis was diagnosed in adults despite a much lower incidence of the illness in 2001 compared to 2000. Thus this pathology should not be underestimated in adults. Considerable medical expenditure might be avoided (cumulative numbers of 172 days in hospital and 82 days of antibiotics in this study), if rapid and accurate diagnostic techniques were available.

Circulation of enteroviruses and persistence of meningitis cases in the winter of 1999-2000.

The seasonal incidence of enterovirus meningitis was analyzed in a prospective study of patients admitted for suspected meningitis from October 1, 1998 to April 30, 2000. In-house reverse transcription-polymerase chain reaction (RT-PCR) in cerebrospinal fluid (CSF) was used irrespective of cytological results. Fifty-two (45.2%) of the 115 patients had positive RT-PCR in CSF, including 44/86 children (51.2%) and 8/29 adults (27.6%). Six of the 52 (11.5%) had no pleocytosis. The numbers of CSF specimens with a predominance of lymphocytes or a predominance of neutrophils were closely similar. In 33 of the positive patients, an enterovirus, mainly echoviruses type 6 (48%) and 30 (24%), was recovered in one or more specimens. Sixteen cases of enteroviral meningitis were observed between November 1999 and March 2000 as against 2 cases between November 1998 and March 1999, showing that the disease persisted through the winter months of 1999-2000. During the same period, 96 enterovirus isolates were recovered from clinical specimens from other patients. The number of isolates was higher in the winter of 1999-2000 (P < 0.01) than in the winter of 1998-1999, indicating that the risk of enterovirus infection increased significantly in winter 1999-2000. Sixteen patients had aseptic meningitis, made a rapid recovery and had an enterovirus in throat swabs and stools (9/16) or in one of the two (7/16). RT-PCR was not requested. Nine patients were admitted during the cold months. The clinical management of both adult and child patients could be improved by year-round use of enterovirus generic RT-PCR.

Prospective analysis of 61 cases of enteroviral meningitis: interest of systematic genome detection in cerebrospinal fluid irrespective of cytologic examination results.

BACKGROUND: Enteroviruses are the most commonly identified cause of viral meningitis. Detection of the enterovirus genome in cerebrospinal fluid (CSF) using reverse-transcription polymerase chain reaction (PCR) has proved to be useful in diagnosis and is more rapid and sensitive than viral cultures. In routine practice, cytologic examination results of CSF are obtained swiftly and PCR indication is performed as a second step. OBJECTIVES: The aim of this study was to determine, by analysis of complete data from CSF results for 61 cases of proven enteroviral meningitis, whether cytologic CSF findings can be used to establish viral etiology and to indicate if PCR assay should be performed. STUDY DESIGN: From a prospective study of children admitted during 1997 for suspected enterovirus meningitis in which PCR and viral cultures of CSF were systematically performed, we selected 61 patients with proven enterovirus meningitis. We compared global white cell count (WCC), relative percentage of lymphocytes/neutrophils, PCR and culture for enterovirus, patient age, and clinical data. RESULTS: 92% of patients (56/61) had positive PCR in CSF and in 48% (29/61) enterovirus was isolated in CSF. Nine patients (14.75%) had WCC<10/mm(3); eight of them had positive PCR and two had positive culture. There were comparable numbers of CSF with a predominance of lymphocytes (n=25) and CSF with a predominance of neutrophils (n=22), and of positive PCR and positive cultures of CSF in the two groups. Results were not influenced by the age of the patients. CONCLUSION: Irrespective of other CSF parameters, it seems difficult to dispense with PCR assay for enterovirus genome detection. It should be introduced as a true rapid routine test. Early reporting of a positive PCR result could result in a considerable saving in health resources.

Nosocomial transmission of echovirus 30: molecular evidence by phylogenetic analysis of the VP1 encoding sequence.

We investigated six cases of enterovirus infection in a neonatal unit. The index patient, a 5-day-old boy, was admitted with aseptic meningitis due to echovirus 30 (E30). Secondary infections with E30 occurred in five babies. Comparison of the complete VP1 sequences showed that the isolates recovered from the index patient and his mother were closely related to those recovered from the five babies with secondary infections, demonstrating a nosocomial transmission of the virus. In the phylogenetic tree reconstructed from the VP1 sequences, the isolates formed a monophyletic cluster related to an E30 strain collected in June 1997 during an outbreak of aseptic meningitis.

Effects of liver transplantation on long-chain polyunsaturated fatty acid status in infants with biliary atresia.

BACKGROUND: The long-chain polyunsaturated fatty acid (LC-PUFA) status of infants with untreated biliary atresia (BA) is known to be poor and is correlated to the severity of the liver disease. Liver transplantation (LT) markedly increases survival of patients with BA but the extent to which this reverses poor LC-PUFA status is not known. METHODS: To explore this question, the erythrocyte (red blood cell, RBC) phospholipid content of eight infants with BA who underwent LT was determined 2 months after an initial portoenterostomy, immediately before LT, and 6 and 12 months after LT. Before LT, all infants were fed a protein hydrolysate formula containing medium-chain triglycerides and essential fatty acids. Afterward, they were fed a normal diet for age. The RBC phospholipid content at each time point was compared with that of 28 age-matched control infants. RESULTS: Just before LT, median RBC phospholipid content of C20:4n-6, C20:5n-3, and C22:6n-3 was 25%, 48%, and 30% lower, respectively, than that observed in age-matched control infants. After LT, the RBC phospholipid content of most fatty acids reached normal values by 6 months. However, that of C20:4n-6 and C22:6n-3 contents remained 5% and 15% lower, respectively, than in normal control infants. Twelve months after LT, C20:4n-6 content remained lower than in normal children, but that of C22:6n-3 did not differ. The ratio of C20:3n-6/C20:4n-6, a reflection of delta-5 desaturase activity, was abnormal compared with normal children before LT (0.17 vs. 0.10, P < 0.009) but normalized by 6 months after LT (0.11 vs. 0.10, not significant). CONCLUSIONS: These data show that the abnormal LC-PUFA status of children with BA improves after LT but is not entirely reversed within a year after surgery. They suggest that the abnormal status before LT may be secondary, in part, to low delta-5 desaturase activity. The extent to which a different pre- and/or post-LT diet can prevent PUFA deficiency and/or hasten recovery of PUFA status remains to be determined.

Genomic variations in echovirus 30 persistent isolates recovered from a chronically infected immunodeficient child and comparison with the reference strain.

Seven sequential isolates of echovirus type 30 (EV30) were recovered over 22 months from a child with severe combined immune deficiency syndrome. The nucleotide sequences of the 5' halves of the genomes (4,400 nucleotides) of the first (S1) and last (S7) isolates were determined and compared with that of the EV30 Bastianni reference strain, also determined in this study. In genome regions P1 and P2, 101 variations were identified between the two isolates. Synonymous differences far outnumbered nonsynonymous differences. Amino acid changes affected both capsid and nonstructural polypeptides (particularly 2B). The VP1 nucleotide sequences of the seven isolates were determined to analyze genome evolution during the chronic infection. In the phylogenetic tree, the seven isolates were directly related to the prototype strain in an individual monophyletic group, strongly suggesting that the chronic infection in the child arose from a single persistent EV30 isolate. Four lineages were observed in the persistent isolates. Isolates S2, S4, S5, and S6 were close relatives of one another, whereas isolates S1 and S3 formed individual lineages. Isolate S7, distantly related to all other isolates, formed the fourth lineage. These findings suggest the quasispecies nature of the genomes of the seven sequential EV30 isolates. Grouping of persistent isolates on the basis of replicative capacities was consistent with phylogenetic relationships. Overall, the results indicate that genetically related EV30 variants with different replicative capacities coexisted in a carrier state, probably in the gastrointestinal tract, during the infection of the child.

An outbreak due to echovirus type 30 in a neonatal unit in France in 1997: usefulness of PCR diagnosis.

Between February and August 1997, 53 patients with enterovirus meningitis were hospitalized in Clermont-Ferrand, France. All but one were children. Echovirus type 30 was involved in 70% of cases with identified serotype. The outbreak ceased on August 8. Two months later, a neonate was admitted to the neonatal unit with an echovirus type 30 meningitis thought to be acquired at delivery. Twenty days later a nosocomial outbreak of echovirus type 30 involving five neonates occurred. Two of them presented with meningitis and two with febrile seizure; One was asymptomatic. The retrospective examination of the maternal sera in a neutralization test, using the index case strain as a source of antigen, showed that none of the neonates was passively immunized before hospitalization. The use of genome detection in cerebrospinal fluid allowed rapid diagnosis and infection was contained by re-inforcing hygiene measures. Prospective examination of stools in the neonatal and paediatric units showed no further occurrences of the disease. No sporadic case was observed in the general population. Hence, nosocomial infections can occur a long time after an outbreak in the general population; rapid diagnosis with molecular tools is useful both for a definite diagnosis in patients already hospitalized, and to act as a rapid alert, even in intervals between seasonal outbreaks.

[The re-emergence in 1997 of rubella infections during pregnancy. 11 cases in Clermont-Ferrand]. / Recrudescence en 1997 des infections rubéoliques pendant la grossesse. 11 cas à Clermont-Ferrand.

OBJECTIVE: Describe the clinical and laboratory features of rubella observed during the first semester of pregnancy in 11 patients in 1997. PATIENTS AND METHODS: Eleven pregnant women, aged 15-30 years, were referred to the Clermont-Ferrand University Hospital for suspected rubella. Four had had at least 1 prior pregnancy, none had been vaccinated. Rubella serology was obtained for all 11 patients and polymerase chain reaction viral amplification was performed on amniotic fluid in 9 cases. RESULTS: The virology laboratory identified 8 cases of primary rubella (2 prior to 12 weeks gestation) and 3 reinfections (1 prior to 12 weeks gestation). Fetal infection was evidenced in I gravida II patient at 17-18 weeks gestation. All pregnancies were continued to term and no case of congenital rubella malformation was observed. However specific IgM assays were performed at birth in 6 of the 11 infants and revealed infection in 3. CONCLUSION: These observations indicate that a local epidemic of rubella occurred in the general population. They illustrate the risk of a rubella epidemic in France and the lack of sufficient vaccination of the young adult population, finally they emphasize that current anti-rubella vaccination programs should be promoted.

Interaction of doxorubicin with ATP: quantification of complexes and effect on its diffusion into DNA-loaded liposomes--implication for ATP-driven transport studies.

Doxorubicin, a drug largely used in chemotherapy, is transported by P-glycoprotein, a protein involved in the multidrug-resistance phenotype. Taking advantage of the doxorubicin fluorescence quenching upon interaction with DNA, a sensitive assay of this active transport can be carried out: quantitative in vitro studies could be achieved with DNA-loaded proteoliposomes, after correction for the doxorubicin passive diffusion through phospholipids. In this paper, we describe experimental conditions that will be relevant to P-glycoprotein studies. Efficient DNA entrapment in preformed liposomes was obtained using the freeze/thawing procedure, and the doxorubicin passive diffusion was quantified in the presence of ATP/Mg2+, the second substrate of P-glycoprotein. The doxorubicin diffusion rate decreases in the presence of ATP, indicating an interaction between doxorubicin and ATP that will hinder any measurement of ATP-driven transport. The interaction between doxorubicin and ATP was studied by fluorescence quenching, octanol/buffer partition coefficient, and diffusion rate into DNA-loaded liposomes. The results give evidence for complex interactions. However, under our experimental conditions, these interactions are only slightly modified in the presence of Mg2+. Since this cation is essential for P-glycoprotein activity, it can be concluded that in these conditions the accurate evaluation of P-glycoprotein-catalyzed doxorubicin transport will be obtained from the Mg2+-sensitive transport into DNA-loaded proteoliposomes.

Comparative sensitivities of Sabin and Mahoney poliovirus type 1 prototype strains and two recent isolates to low concentrations of glutaraldehyde.

Significant intratypic differences in the glutaraldehyde (GTA) sensitivity of echovirus isolates have been shown. While exploring ways to optimize the study of GTA sensitivity of enteroviruses, we also observed intratypic differences in poliovirus type 1 isolates collected in France. A suspension procedure was used for assessing the virucidal effect of GTA at low concentrations (< or = 0.10%) against purified viruses. Two recent isolates of poliovirus type 1 tested were first fully characterized by the PCR restriction fragment length polymorphism (RFLP) test. The RFLP pattern of clinical isolate 5617 was similar to that of poliovirus type 1 LS-c, 2ab (Sabin strain), confirming the vaccine origin of strain 5617. The RFLP pattern of strain 5915 recovered from sewage was different from that of the Mahoney strain, suggesting a genetic variation in this wild isolate. We then analyzed under the same controlled conditions the GTA sensitivities of both isolates and their respective prototype strains. The wild Mahoney and 5915 strains exhibited significantly lower sensitivities to GTA than did the vaccine Sabin and 5617 strains. The inactivation rates of clinical isolates 5617 and 5915 were very similar to those of their corresponding reference Sabin and Mahoney strains. Both the conformational structure of the capsid of each strain and the amino acid constitution of structural polypeptides could be involved in the variations observed. The relevance of our comparative sensitivity studies to standardization of virucidal tests is discussed.