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.

[Diagnosis and epidemiological surveillance of influenza and respiratory syncytial virus infections: interest of multiplex PCR]. / Diagnostic et surveillance épidémiologique des infections grippales et à virus respiratoire syncytial: intérêt de la PCR multiplex.

OBJECTIVE: Respiratory infections require a rapid etiological diagnosis for efficient management of cases. We evaluated multiplex PCR used for the diagnosis and the epidemiological surveillance of influenza and respiratory syncytial virus (RSV) infections. PATIENTS AND METHODS: Our study included 278 patients (mean age: 37.2+/-22.9 years) with flu or flu-like syndromes, consulting physicians affiliated with the GROG Poitou-Charentes or hospitalized in the Poitiers teaching hospital. A multiplex PCR detecting A(H3), A(H1) and B influenza viruses, and RSV A and B, was performed with both a direct examination by immunofluorescence and cell-culture. RESULTS: We diagnosed a viral infection in 139 (50.0%) patients: 99 cases of influenza A(H3), 2 cases of influenza A(H1), 28 cases of influenza B and 11 cases of RSV infections. The diagnosis yield in GROG patients (52.3%) was significantly higher than that observed in hospitalized patients (34.5%) (P=0.04). All techniques were correlated in 61% of cases. The multiplex PCR yielded 22.3% more positive samples compared to the conventional techniques. All positive samples by conventional techniques were also positive by multiplex PCR. We observed a perfect correlation between viral types and subtypes determined by PCR and cell-culture. CONCLUSION: Multiplex PCR is a sensitive technique allowing an efficient and rapid diagnosis of respiratory infections due to influenza and RSV.

Low serum selenium concentrations in French patients with measles.

OBJECTIVE: Selenium deficiency adversely affects the clinical outcome of measles in the tropics. In developed countries, serum selenium level has never been investigated during acute measles. The aim of this study was to determine serum selenium concentrations in French patients with acute measles and to seek correlations with clinical and virological findings. PATIENTS AND METHODS: We studied serum selenium concentrations in 94 French patients with acute measles and in 99 healthy controls matched for age and sex. RESULTS: The mean of selenium concentration was significantly lower in the patients than in the controls (46.4±14.1µg/L versus 86.5±13.9µg/L, P<0.0001). In the patients, selenium concentrations were not associated with age, sex, vaccination status, clinical signs or specific antibody responses. Selenium levels did not differ significantly between patients with uncomplicated measles (45.8±14.2µg/L) and patients with complications (52.7±13.2µg/L) (P=0.15). CONCLUSION: Acute measles is associated with significant reduction of selenium level that did not seem to negatively affect the course of the disease suggesting compensating mechanisms in patients from developed countries against the disease.

Clinical severity and molecular characteristics of circulating and emerging rotaviruses in young children attending hospital emergency departments in France.

Group A rotavirus (RVA) is the leading cause of acute gastroenteritis in young children worldwide. A prospective surveillance network has been set up to investigate the virological and clinical features of RVA infections and to detect the emergence of potentially epidemic strains in France. From 2009 to 2014, RVA-positive stool samples were collected from 4800 children <5 years old attending the paediatric emergency units of 16 large hospitals. Rotaviruses were then genotyped by RT-PCR with regard to their outer capsid proteins VP4 and VP7. Genotyping of 4708 RVA showed that G1P[8] strains (62.2%) were predominant. The incidence of G9P[8] (11.5%), G3P[8] (10.4%) and G2P[4] (6.6%) strains varied considerably, whereas G4P[8] (2.7%) strains were circulating mostly locally. Of note, G12P[8] (1.6%) strains emerged during the seasons 2011-12 and 2012-13 with 4.1% and 3.0% prevalence, respectively. Overall, 40 possible zoonotic reassortants, such as G6 (33.3%) and G8 (15.4%) strains, were detected, and were mostly associated with P[6] (67.5%). Analysis of clinical records of 624 hospitalized children and severity scores from 282 of them showed no difference in clinical manifestations or severity in relation to the genotype. The relative stability of RVA genotypes currently co-circulating and the large predominance of P[8] type strains may ensure vaccine effectiveness in France. The surveillance will continue to monitor the emergence of new reassortants that might not respond to current vaccines, all the more so as all genotypes can cause severe infections in infants.

ORL-1 and mu opioid receptor antisera label different fibers in areas involved in pain processing.

Mu opioid receptors (MOR) mediate the analgesic effects of opioid drugs such as morphine. The opioid receptor-like (ORL-1) receptor is structurally related to opioid receptors and the ORL-1 receptor agonist, orphanin FQ/nociceptin, induces analgesia at the spinal level, but appears to recruit different circuitry than that used by mu opioids. When administered intracerebroventricularly, orphanin FQ/nociceptin produces hyperalgesia and/or reverses opioid analgesia. The functionally distinct actions elicited by MOR and ORL-1 receptors, which activate similar intracellular signaling systems and show similar regional distributions, could be explained by their differential cellular localization. By using double label immunohistochemistry and confocal microscopy, the present study investigates the distribution of MOR and ORL-1 receptors in regions of the rat nervous system that are involved with nociceptive processing. In general co-localization of MOR and ORL-1 receptor immunoreactivity was not observed in either perikarya or neuropil in the dorsal root ganglia, nor in the Lissauer's tract and superficial laminae of the spinal cord. Likewise, there was no evidence for co-localization of these receptors within the periaqueductal gray, the nucleus raphe magnus, the gigantocellular reticular nucleus, and the nucleus of the solitary tract. These observations indicate that MOR and ORL-1 receptors are expressed predominantly on different fiber systems in these regions. This differential distribution is consistent with the distinct pharmacology of ORL-1 and MOR receptor agonists and suggests that the antisera to MOR and ORL-1 receptors may provide useful markers for further investigations of analgesic and counteranalgesic pathways modulating pain perception.

In vitro characterization of a biologically active molecular clone of HIV-2NIH-Z containing a nef deletion and expressing a full-length transmembrane protein.

We have previously described the cloning and sequencing of a novel stain of human immunodeficiency virus type 2 (HIV-2) called HIV-2NIH-Z. A plasmid clone, pHIV2Z, containing the full-length provirus has now been constructed, and virus particles have been obtained upon transfection into COS-1 and H-9 cells. These particles can infect a number of T-cell lines and exert a cytopathic effect on fresh human and macaque peripheral blood lymphocytes. The cloned virus is biologically and morphologically indistinguishable from its parental uncloned strain as shown by restriction enzyme analysis, electron microscopy, and kinetics of infection. However, as shown by radioimmunoprecipitation assays, the cloned virus-infected cells express a full-length gp41 protein as predicted by the nucleotide sequence, whereas the wild-type parental strain expresses a truncated gp33 protein. Both the parental strain and the cloned virus possess a deletion encompassing the end of the nef gene within the U3 region which apparently does not affect their in vitro cytopathic and replicative capacities.

Polymerase chain reaction amplified HTLV-I, HIV-1 and HIV-2 DNA fragments in subjects with mixed retroviral infections.

Serum and peripheral blood mononuclear cells from eight patients from the Ivory Coast with positive screening test results for retroviral infections were studied by serology (ELISA, Western blot (WB), synthetic peptide test), cell co-culture, and polymerase chain reaction (PCR). Two HIV-2 infections with indeterminate interpretation on HIV-1 WB were detected, two were clear dual HIV-1/HIV-2 infections, three were ambiguous mixed HIV-1/HIV-2 infections, and one was a triple retroviral infection by HTLV-I, HIV-1 and HIV-2. Four slow/low HIV-1 strains were isolated at the expense of HTLV-I and HIV-2 strains. The ELISA tests were found to be very sensitive. Indeterminate WB interpretations were frequent (HTLV-I, four; HIV-1, three; HIV-2, two). PCR provided clear evidence of multiple retroviral infections in three cases and enabled interpretation of indeterminate WB samples in three cases. One sample presented a puzzling pattern with positive PCR results for HIV-1 and HIV-2 associated with negative or indeterminate serological results. Thus, our data emphasise the need to analyse serological as well as virological markers to gain better insight on mixed retroviral infections, especially in endemic areas such as West Africa.

Variable stringency hybridization of polymerase chain reaction amplified HIV-1 DNA fragments.

DNA isolated from the peripheral blood mononuclear cells of HIV-1 seropositive individuals was used for polymerase chain reaction (PCR) amplification of gag and envelope regions. Eight aliquots of the amplified DNA fragments have been subjected to Southern/dot blot analysis, hybridizing with 32P-labelled-BH10 (HIV-1 strain IIIB) at low stringency. After the filters had been autoradiographed, they were cut so that each hybridized band/dot could be subject to variable stringency washing using various ionic concentrations at a fixed temperature. The filter was reconstructed so that the effect of the variable stringency wash might be visualized following a second exposure to Kodak film. The level of activity for each band/dot was measured by counting the 32P or by densitometry analysis of the photographic record. The results allow a plot to be made of the decrease in bound radioactivity against ionic strength. By comparison with a standard curve obtained for HIV-1 strain IIIB amplified fragments subject to similar hybridization and analysis, an estimation of the degree of nucleotide mismatch relative to the BH10 DNA probe can be obtained. The technique provides a rapid means of characterizing PCR amplified fragments.

Unclassifiable high grade malignant T-cell lymphoma with clonal evolution.

We report a case of leukemic malignant T-cell lymphoma with mixed small and large cells. The small cells displayed a mature CD8-positive phenotype, a diploid DNA distribution by cell cycle analysis, and structural karyotypic abnormalities. Large cells were near triploid, showed additional structural cytogenetic abnormalities and a more immature membrane phenotype without CD8 expression. Altogether, these data provide suggestive evidence for a clonal evolution from a mature small cell T-cell lymphoma to a more immature large cell proliferation.

Evaluation of closed sterile prefilled humidification.

A closed sterile prefilled humidifier ('Aquapak 310') and a multiple-use humidifier ('Nebal 2') were evaluated in hospital departments to determine their susceptibility to bacterial contamination and cost. No bacterial contamination was found in the 389 samples of 'Aquapak 310' water. However, 54/164 (32.9%) samples of 'Nebal 2' water were found to be contaminated. Pseudomonas aeruginosa was the bacterium most often isolated. The cost analysis was highly influenced by the average use time. In the haemodialysis and respiratory medicine departments the average use times for the 'Aquapak 310' +/- SD were 61.6 +/- 36.2 days and 4.1 +/- 1.7 days, respectively. Using the 'Aquapak 310' system, there was a 51% financial saving in the haemodialysis department but a 2% loss in the respiratory medicine department. In these two departments we found a similar cost saving as far as staff time was concerned (88% vs. 89%). The major difference came from the cost of consumables: 26% saving in the haemodialysis department vs. 70% loss in the respiratory medicine department. Use of the prefilled sterile humidifiers represents a three-fold benefit, a lower infection risk for the patient, an important financial saving in the haemodialysis department and a decreased staff work load.

Seroprevalences of Toxoplasma, malaria, rubella, cytomegalovirus, HIV and treponemal infections among pregnant women in Cotonou, Republic of Benin.

Seroprevalences for toxoplasmosis, malaria, rubella, cytomegalovirus, HIV and treponemal infections were evaluated among 211 pregnant women residing in the Cotonou area, Republic of Benin. One hundred and thirteen women (53.6%) had toxoplasma antibodies, 185 (87.7%) malaria antibodies and 181 (85.8%) rubella antibodies. Among the 205 (97.2%) women with cytomegalovirus antibodies, 6 presented recent or current infection. No HIV seropositivity was detected. Five (2.4%) of these women had a positive treponematosis serology corresponding to previous infection or reinfection. These results were compared with previous studies conducted in Africa. Routine serological screening should be recommended in young age and in pregnancy for rubella, only in pregnant women for HIV and toxoplasma infections, in order to control their possible consequences on women and newborns.

Maternal Herpes simplex virus type 2 encephalitis following Cesarean section.

Herpes simplex virus type 2 (HSV-2) encephalitis is rare especially during pregnancy. In immunocompetent patients, HSV-2 meningitis (contrary to HSV-1 meningitis) is usually mild, without encephalitis. We report a rare case of maternal HSV-2 encephalitis following Cesarean section. The woman had no symptomatic genital lesion, and the infant was not infected. The route of meningeal infection (neuronal or hematogenous) is discussed.

Genital recurrent infection occurring 6 months after meningitis due to the same herpes simplex virus type 2 (HSV-2) strain evidence by restriction endonuclease analysis.

Herpes simplex virus type 2 (HSV-2) is more often sexually transmitted and associated with genital recurrent infection. However, HSV-2 neurological manifestations such as meningitis were already reported. We describe a case of meningitis due to HSV-2, preceded by signs suggesting a common cystitis, in a woman with no history of primary or recurrent genital infection. Six months later genital herpetic lesions occurred. One HSV-2 strain was obtained from cerebrospinal fluid (CSF) and another from genital lesions. The molecular comparative analysis using restriction endonuclease digestion patterns showed the similarity of the two strains. Our report illustrates that HSV-2 infections are underdiagnosed and that molecular techniques can be of value in clarifying the physiopathology of HSV diseases.

[Evaluation of ELISA using A60 antigen from Mycobacterium bovis BCG: specific IGG and IGM in mycobacterial infections]. / Evaluation d'un ELISA utilisant l'antigène A60 de Mycobacterium bovis BCG: IgG et IgM spécifiques au cours des infections mycobactériennes.

The authors have evaluated an ELISA (A60-Tb, Anda biologicals) allowing the detection of specific IgG and IgM against A60 antigen from Mycobacterium bovis BCG during mycobacterial infections. This study included sera from 110 african subjects and from 71 French subjects distributed in 4 clinical groups: 55 tuberculous patients (I), 41 leprous patients (II), 33 pneumopathies (III) and 52 healthy subjects (IV). Serological results were compared taking as reference for the diagnosis of tuberculosis the positivity of culture and/or that of a direct examination, and for leprosy the positivity of a direct examination associated either with a Mitsuda's reaction or with an histopathological examination. IgG were found to be more discriminative than IgM. Considering together the results of groups I and II, the authors found a sensitivity of 95.8 p. cent and a specificity of 75.3 p. cent with threshold of 200 U/ml for specific IgG. Anti-A60 antigen antibodies obtained for groups I and II were significantly higher (IgG: p less than 0.0001; IgM: p less than 0.001) than those observed in other groups. African subjects presented IgG titers higher than those obtained by French subjects (p less than 0.0001). IgM response was more frequent among group II (97.6 p. cent) than group I (21.8 p. cent). However, IgG (26.9 p. cent) and IgM titers (30.8 p. cent) were detected among group IV. This test would allow a control of therapeutic efficacy with an additional interest for classifying borderline forms of leprosy.

[Nosocomial infections due to rotavirus and respiratory syncytial virus in pediatric wards: a 2-year study]. / Infections nosocomiales à rotavirus et à virus respiratoire syncytial en milieu pédiatrique: étude sur une période de 2 ans.

Rotavirus and respiratory syncytial virus (RSV) infections represent up to 30% of the totality of nosocomial infections in paediatric wards. We studied the importance of these infections in the paediatric wards of the University Hospital Center of Poitiers, France, from October 1996 to September 1998. We defined as nosocomial an infection acquired after 3 days of hospitalization for rotavirus and after 7 days for RSV. The 274 cases of children presenting rotavirus gastroenteritis or RSV infection within this period were studied. Rotavirus was detected in stools by using an agglutination test and RSV was diagnosed in nasopharyngeal aspirations by direct examination with an immunofluorescence assay (IFA), cell culture and serotyping with IFA. We noted 50 rotavirus and 224 RSV infections, with a first epidemic of RSV subgroup B (49.5%) and a second epidemic of subgroup A (44.9%). 19 (38%) were rotavirus nosocomial infections and 5 (2.2%) were RSV nosocomial infections. The majority of the nosocomial infections occurred before the age of one year and particularly before the age of 6 months (42.2% for rotavirus, 60% for RSV). In comparison to community-acquired infections, children with rotavirus nosocomial infections were younger (9 months versus 12.5 months) which was the opposite for RSV nosocomial infections (10.8 months versus 6.5 months). The sex-ratio of children with community-acquired infections was 2.1 that was not reported in nosocomial infections. The length of stay in hospital was always longer in nosocomial infections (11.7 days versus 3.6 days for rotavirus; 38.8 days versus 4.8 days for RSV). Diarrhea (p = 0.007) and vomiting (p = 0.013) for enteric infections and wheezing (p = 0.02) for respiratory infections were more often observed in community-acquired infections. This study emphasizes the frequency and the consequences of rotavirus and RSV nosocomial infections in paediatric wards and the importance of the hygienic rules to prevent these infections.

[Rhinovirus infections in hospitalized children: a 3-year study]. / Infections à rhinovirus chez des enfants hospitalisés: étude sur une période de 3 ans.

BACKGROUND: Rhinoviruses (RH) are responsible for acute respiratory illnesses, mainly in the upper respiratory tract. POPULATION AND METHODS: 3,152 children aged under 16 years, admitted to the Paediatrics department of the University Hospital Centre of Poitiers from January 1, 1993 to December 31, 1995 with ear, nose and throat (ENT) and/or respiratory symptoms were systematically investigated. One hundred and forty-five RH strains were isolated from nasopharyngeal secretions of 87 boys and 58 girls (mean age: 20.3 months). Among these, 92 (63.4%) were less than 1 year of age. Bacteriological investigations were done for 29 patients when a concomitant bacterial infection was suspected. RESULTS: RH infection rate was maximum before 1 year of age (median age: 6.5 months) and decreased with age. RH were isolated throughout the 3 years, with a first peak from February to April, and a second one in autumn. The main symptoms were sibilants (27.6%) and cough (24.1%). Sibilants were more frequently associated in children under 12 months of age (P = 0.01). Sometimes, ophthalmologic or digestive symptoms were present. Three children with respiratory distress were transferred to the reanimation ward. In addition, a RH strain was isolated from a child who died of sudden infant death. Thirty-four children (23.4%) were co-infected by one or several viruses; the most frequently detected were the respiratory syncytial virus (41.2%) and the adenoviruses (35.3%). Twenty-nine children were infected by two viruses and five by three. Associated bacterial infections were diagnosed in 23 children, especially conjunctivitis due to Haemophilus influenzae (21.7%). Among these children, eight had a multiple viral infection. CONCLUSION: RH have a limited pathogenicity but can be associated with serious illnesses among infants and children.

[Q fever and fetal death in utero. Two cases]. / Fièvre Q et mort foetale in utero. Deux observations.

Some cases of late abortion occurring after a Coxiella burneti infection, more often with a chronic evolution, have already been mentioned in the literature. We reported here two cases of early abortion, contemporaneous of an acute infection due to C. burneti. Two patients, after a contact, before and at the beginning of the pregnancy, with an animal susceptible to contaminate human beings by C. burneti, presented no clinical symptom characteristic of Q fever. The fetal death for the two cases was found out at the 17th week of amenorrhoea. All the investigations in order to search for an abortion etiology remained negative. Only, the specific serologies showed an acute infection due to C. burneti.

[Epidemic of viral gastroenteritis in an elderly community]. / Epidémie de gastro-entérite virale dans une collectivité de sujets âgés.

Rotaviruses and adenoviruses are the main aetiologic pathogens of gastroenteritis in infants. Adults may also be concerned with usually mild clinical symptoms. An epidemic of viral gastroenteritis occurred in a long-term hospitalization ward from July 25 to August 21, 1991: 101 patients. This study involved 26 females and 6 males with a mean age of 86 years (range 70-101 years), presenting clinical symptoms of gastroenteritis. The clinical attack rate was 27.7%. Outcome was favourable for the majority of patients who recovered in 4-5 days. The main clinical signs were diarrhoea (90.6%), hyperthermia (18.8%) and vomiting (18.8%). Virology investigations gave the diagnosis of 8 rotavirus infections, 6 adenovirus infections and 2 rotavirus and adenovirus mixed infections. Among staff members, 3 rotavirus and one adenovirus infections were diagnosed. The precise origin of the epidemic could not be determined from rotavirus electropherotypes obtained from stools of elderly and paediatric patients hospitalized during the same period. This outbreak recalls the viral involvement in diarrhoeal episodes of elderly people. These episodes of viral gastroenteritis are responsible for high morbidity in the elderly and may upset a precarious physiological state.

[Anti-BCG titers of healthy and tuberculous Africans. Use of the indirect immunoperoxidase test]. / Titrage d'anticorps anti-BCG chez des africains sains et tuberculeux. Utilisation d'un test indirect à l'immuno-peroxydase.

This study was intended to assess the presence of antibodies to BCG: immunoglobulin G (IgG) and immunoglobulin M (IgM) in three homogenous african populations: 76 tuberculous patients, 55 adult healthy subjects and 46 newborn. We have used an indirect immuno-peroxidase reaction. Tuberculous patients were clearly distinguished from the other two groups by raised IgG titres, associated with severe clinical and radiological features. In the control population, the IgG anti-BCG were also present, but to a lesser degree. The IgM anti-BCG were seen in high titres (greater than 20 in this study) in healthy adults and patients. The two populations differed significantly (p less than 0.001), nevertheless some healthy adults achieved IgM titres comparable with tuberculous patients. This simple test could be an interesting contributory factor in cases of diagnostic difficult and enables a serological assessment of patients having BCG therapy.

[Resistance of Mycobacterium tuberculosis. An 8-year survey in the Poitiers area]. / Résistance de Mycobacterium tuberculosis. Enquête sur 8 ans en région poitevine.

We have studied in Poitiers area from 1977 to 1984 the resistance of 853 Mycobacterium tuberculosis strains to the main anti-tuberculosis drugs. The overall rate of drug resistance was showed to be steady over the years while the primary resistance rate has decreased. The only one drug resistance has concerned para-aminosalicylic acid, streptomycin and isoniazid. Foreigners, most of them Asian people or North-African people, often bear resistant tubercle bacilli (29,03%) compared with French people (9,29%). We encountered drug resistance phenomena essentially among less than 60 years old patients. Drug susceptibility tests remain indispensable for a good epidemiologic supervision at the time of relapses and among patients possibly infected with multiresistant germs.