Interest of human papillomavirus DNA quantification and genotyping in paired cervical and urine samples to detect cervical lesions.

BACKGROUND: Cervical cancer is caused by persistent infection with high-risk human papillomavirus (HR-HPV). Conventional human papillomavirus (HPV) testing requires cervical sampling. However, vaginal and urine self-sampling methods are more acceptable for patients and result in increased participation when they are available in screening programs. In this context, we have developed a non-invasive screening method via the detection of HPV DNA in urine samples. PURPOSE: To compare HPV viral loads and genotypes in paired cervical and urine samples, and to assess correlation between virological and cytological results in women seeking gynecological consultation. METHODS: Paired urine and cervical specimens were collected and analyzed from 230 of 245 women participating in the previously described prospective PapU study. HPV DNA detection and quantification were performed using a real-time PCR method with short fragment PCR primers. Genotyping was carried out using the INNO-LiPA HPV genotyping assay. RESULTS: The prevalence of HPV in the 230 paired urine and cervical smear samples was 42 and 49 %, respectively. Overall agreement for HPV positivity and negativity between the paired samples was 90 % (κ = 0.80). High HPV viral load in both cervical and urine samples was associated with cytological abnormalities. HPV-positive women were mostly infected with HR-HPV types. The agreement between high- and low-risk HPV (LR-HPV) detection in both samples was 97 % (κ = 0.95 for HR-HPV and κ = 0.97 for LR-HPV). CONCLUSIONS: High concordance rates for HPV-DNA quantification and high/low-risk HPV genotyping in paired urine/cervical samples suggest that urinary HPV DNA testing could be useful for cervical lesion screening.

Surveillance network for herpes simplex virus resistance to antiviral drugs: 3-year follow-up.

Herpes simplex virus (HSV) infections are very common in the general population and among immunocompromised patients. Acyclovir (ACV) is an effective treatment which is widely used. We deemed it essential to conduct a wide and coordinated survey of the emergence of ACV-resistant HSV strains. We have formed a network of 15 virology laboratories which have isolated and identified, between May 1999 and April 2002, HSV type 1 (HSV-1) and HSV-2 strains among hospitalized subjects. The sensitivity of each isolate to ACV was evaluated by a colorimetric test (C. Danve, F. Morfin, D. Thouvenot, and M. Aymard, J. Virol. Methods 105:207-217, 2002). During this study, 3900 isolated strains among 3357 patients were collected; 55% of the patients were immunocompetent. Only six immunocompetent patients excreted ACV-resistant HSV strains (0.32%), including one female patient not treated with ACV who was infected primary by an ACV-resistant strain. Among the 54 immunocompromised patients from whom ACV-resistant HSV strains were isolated (3.5%), the bone marrow transplantation patients showed the highest prevalence of resistance (10.9%), whereas among patients infected by human immunodeficiency virus, the prevalence was 4.2%. In 38% of the cases, the patients who excreted the ACV-resistant strains were treated with foscarnet (PFA), and 61% of them developed resistance to PFA. The collection of a large number of isolates enabled an evaluation of the prevalence of resistance of HSV strains to antiviral drugs to be made. This prevalence has remained stable over the last 10 years, as much among immunocompetent patients as among immunocompromised patients.

Neonatal nosocomial respiratory infection with coronavirus: a prospective study in a neonatal intensive care unit.

The aim of this prospective study was to evaluate the incidence of viral respiratory infection in hospitalized premature newborn infants and to assess the role of coronaviruses. All hospitalized premature infants with a gestational age less than or equal to 32 weeks were included. Tracheal or nasopharyngal specimens were studied by immunofluorescence for coronaviruses, respiratory syncytial virus, adenoviruses, influenza and parainfluenza viruses. Forty premature infants were included; 13 samples were positive in 10 newborns (coronaviruses n = 10; influenza 1 n = 2; adenovirus n = 1). None was positive at admission. All premature infants infected with coronaviruses had symptoms of bradycardia, apnea, hypoxemia, fever or abdominal distension. Chest X-ray revealed diffuse infiltrates in two cases. However, no significant difference was observed between infected and non-infected premature infants for gestational age, birth weight, duration of ventilation, age at discharge, incidence of apnea or bradycardia. Nosocomial respiratory tract infection with coronaviruses appears to be frequent. The clinical consequences should be evaluated in a larger population.

Detection of antibodies against human papillomavirus (HPV) type 16 virions by enzyme-linked immunosorbent assay using recombinant HPV 16 L1 capsids produced by recombinant baculovirus.

The L1 major capsid protein of human papillomavirus type 16 (HPV-16) was expressed in Sf-21 insect cells with a recombinant baculovirus. Virus-like particles obtained were purified and used to develop an enzyme-linked immunosorbent assay for detection of anti-HPV-16 antibodies in sera from 76 women with evidence of genital HPV infection and 79 controls. HPV-16-infected individuals developed antibodies directed at HPV-16 virions since reactivity against recombinant HPV-16L1 capsids was observed in 50% of them compared with only 6% in the general adult population. However, some cross-reactivities with sera from women infected with others HPV types were observed.

Detection of antibodies to L1, L2, and E4 proteins of human papillomavirus types 6, 11, and 16 by ELISA using synthetic peptides.

Antibodies against eight synthetic peptides spanning different epitopes located on L1, L2, and E4 proteins of human papillomavirus (HPV) types 16, 6, and 11 were examined in sera from 73 women infected by HPV and from 139 healthy controls. Only three of these peptides were reactive. Two located on proteins L2 and E4 of HPV 16 seem type specific since antibodies to these peptides were detected, respectively, in 21% and 15% of the HPV 16 infected patients and in 2.5% and none of women infected by other HPVs. The third peptide located on the L1 protein of HPV 6 bears a common epitope since antibodies to this peptide were detected not only in 85% of women infected by HPV 6 or 11, but also in 82% of women infected by other HPVs, and in 74% and 71% of the control groups (10-12-year-old children and adults, respectively). In conclusion, none of the peptides investigated seems useful to develop ELISAs for serological diagnosis of HPV infection.

[Pathogen role of coronavirus in pediatric intensive care: retrospective analysis of 19 positive samples with indirect immunofluorescence]. / Rôle pathogène des coronavirus en réanimation pédiatrique: analyse rétrospective de 19 prélèvements positifs en immunofluorescence indirecte.

BACKGROUND: Coronaviruses (CV) are ARN-containing agents that are responsible for mild upper respiratory tract infections in adults and children. Their pathogenicity in neonates is not wellknown. POPULATION AND METHODS: Eighty five samples of tracheal or nasopharyngeal secretions were obtained from January to October 1991 from 53 children (29 less than 1 month of age; 19 from 1 month to 1 year; five older than 1 year). They were examined for respiratory syncytial virus, adenovirus, myxovirus influenza and parainfluenza and coronavirus by immunofluorescence (IF). RESULTS: Nineteen samples from nine newborns and three infants were positive for coronavirus. The mean birth weight of the nine neonates was 2100 +/- 840 g; their mean gestational age was 34 +/- 5 weeks and their mean age at diagnosis was 21 +/- 9 days. Apnea and bradycardia were the main symptoms in this group. Blood C-reactive protein was not elevated and blood cultures were sterile. One infant was admitted for near-miss; the two others were admitted at birth because they suffered from chronic lung disease (pulmonary hypoplasia and bronchopulmonary dysplasia). An acute episode of pulmonary deterioration occurred at the time of coronavirus infection. Both died one at 4 months and the other at 10 months. CONCLUSION: Coronaviruses seem to be responsible for respiratory tract infections in hospitalized neonates and chronically ventilated infants.

[2 outbreaks of adenovirus 8 keratoconjunctivitis. Clinical, epidemiological and preventive approach]. / Deux vagues épidémiques de kérato-conjonctivite dues à l'adénovirus 8. Approche clinique, épidémiologique et prophylactique.

Adenovirus 8 represents the main agent of Epidemic Keratoconjunctivitis (EKC) and outbreaks mainly occur in hospital wards, colleges and metallurgy industry. Adenovirus 8 conjunctivitis are generally severe lasting two to three weeks, and keratitis with persistent infiltrates may be observed for more than a year. There is no actual treatment of EKC and therefore preventive measures appear as essential to avoid and/or limit such outbreaks in hospital wards. We have observed two consecutive outbreaks of Adenovirus 8 EKC in the Ophthalmology Department of the University Hospital in Brest, France. One outbreak lasted from December 1st, 1983, to March 20, 1984, and the second from July 13, 1984, to November 6, 1984. All patients were selected on the basis of positive virus isolation and they were divided into three epidemiological groups: Group 1 concerned patients who came for EKC treatment at the Ophthalmology Department for the first time; Group 2 resembled either infected staff members or patients which came for EKC and which had already been at the Ophthalmology Department for less than a month with another disease; Group 3 corresponded to hospitalized patients which had contracted EKC during their stay in the ward. Virus isolations were attempted at the virus laboratory in Brest (Pr Chastel) using human diploid fibroblastic cells, MRC 5 strain (Bio-Mérieux, France) and virus isolates were identified by neutralisation tests.(ABSTRACT TRUNCATED AT 250 WORDS)