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ABSTRACT- Using direct immunofluorescence, a nuclear antigen was found in liver of chronic hepatitis patients with circulating NANBe Ag or anti-NANBe, and selected sera from either group were used as source of conjugates. The new Ag/Ab system was designated NANBc Ag and anti-NANBc since it behaved like the core Ag of HBV. N ANBc Ag was detected in coded frozen liver biopsies from patients with chronic persistent 15/25 (60 %) or active 27/50 (54 %) hepatitis and cryptogenic cirrhosis 16/30 (53.3%) devoid of HBV markers. Only 2/30 alcoholic cirrhosis cases (7%) used as controls were positive (p≤0.001). The homologous anti-NANBc antibody was always detectable by indirect immunofluorescence in the patients' serum when N ANBc Ag was found in the liver. It was also found in 11/135 (8%) additional cases without any other NANB marker. A correlation was observed between coded detection of the NANBc Ag/Ab system by immunofluorescence and demonstration of NANBe Ag or anti-NANBe by immunodiffusion. In acute post-transfusion NANB hepatitis, anti-NANBc was first detectable 14 days after transfusion and persisted as long as ALT remained elevated, or longer. IgM anti-NANBc present at onset became associated with an increasing proportion of IgG after the 28th day. The prevalence of anti-NANBc in sporadic NANB hepatitis (11/50 = 22%) was significantly lower (p≤0.001) than in cases with parenteral exposure such as post-transfusion, occupational or drug addict hepatitis (47/72 = 65%). Immunofluorescent tests for NANBc Ag and Ab are promising assays for the serological diagnosis of NANB hepatitis. [ABSTRACT FROM AUTHOR]

Objectives: A campaign designed to enhance screening for hepatitis C virus infection (HCV) by private practitioners was conducted in the Lyon area from April 2000 to March 2001. The target population for screening was composed of individuals with active/previous intravenous drug abuse habits or had received blood products before 1991 or who presented elevated serum transaminase levels. The objectives of the present study were to validate the campaign's organization and its impact on screening practices., Methods: The impact of the campaign was estimated from the number of prescribed and performed HCV screening and confirmation tests, the number of physicians who prescribed HCV screening tests, and the number of HCV screening tests prescribed by each physician. Indicators/practices observed during the 12-month campaign were compared with the same indicators/practices observed during the 12-month period preceding the campaign., Results: During the campaign, 15952 HCV serology tests were prescribed by 1798 of the 3052 physicians (58.9%) practising in the Lyon area. Among physicians who prescribed serology, 11.4% of the general practitioners and 10.9% of the specialists prescribed HCV serology tests for the first time during the campaign. Prescriptions for HCV serology increased significantly between the two calendar periods (+15.6%, epsilon = 17.7), similarly for general practitioners and specialists. The number of general practitioners who prescribed more than five serology tests per year increased 15.3% while the number of general practitioners who did not prescribe serology tests decreased 9.7% (P = 0.02). More HCV-positive patients were detected during the campaign (276 patients) than during the previous period (231 patients) [epsilon = 2.8]., Conclusion: When provided with pertinent information and training adapted to their medical practice, private practitioners become actively involved in screening for HCV infection.

Objective: A screening campaign requested by the French Health Insurance Fund was performed in the Lyon area to access the prevalence of anti-hepatitis C virus in patients from 271 general practitioners and to observe the follow-up of the patients with had positive serology. A total of 101 patients had already had hepatitis C virus antibodies and 30 patients had newly detected antibodies against hepatitis C virus. All subject received appropriate medical care. The aim of this study was to analyse medical follow-up., Methods: Follow-up was performed for one year by the patient's general practitioner or a specialist of liver disease. Clinical, biological, histological and virological data were collected and treatment was proposed when appropriate. Information was gathered anonymously by phone., Results: Follow-up was regular for 28 patients (93%). During this follow-up, ALAT levels remained within the normal limit for 13 patients (43%) and 6 of them had undetectable viral RNA by PCR. In the other patients (50%), a liver biopsy was affered to 11 patients (40%), and performed in 9 (30%). Treatment was started in 8 patients., Conclusion: The results of this study on medical follow-up are satisfying. The general practitioner plays a key role in the follow-up of patients infected by hepatitis C virus.

A study on the screening practices for hepatitis C was carried out among a sample of 786 general practitioners in the Lyon region. The sample comprised three groups: 272 doctors who had participated in a study of this screening, 242 doctors who had not responded to the request for participation, and 272 doctors who had refused to participate. Overall, 88.5% of the doctors surveyed had prescribed at least one HCV serology in the last 12 months, 82% had at least one HCV positive patient and of them, 59.3% had partly or completely screened these patients. The only differences between the three groups are related either to their belonging to an HIV or drug addict care network, or to a particular medical practice (study of allergies, sexology, nutrition, etc.). In fact, the first prescribe more among groups at risk and the second prescribe less and have fewer HCV positive patients.

The aim of this study was to evaluate the Chiron branched DNA (bDNA) assay for detection of serum hepatitis B virus (HBV) DNA in patients with chronic hepatitis B lacking hepatitis B e antigen (HBeAg) and undergoing interferon (IFN) therapy. Results obtained with the bDNA assay were compared with those obtained using the Abbott liquid hybridization (LH) assay and the polymerase chain reaction (PCR). Serial samples (274) from 34 patients were analysed. Analysis of variance results indicated that bDNA values were more significantly correlated than LH values with both PCR positive/negative results (probability of artifact (Prob > F) = 0.7 and 0.09 for LH and bDNA assays, respectively) and presence/absence of precore mutations (Prob > F = 0.21 and 0.001 for LH and bDNA assays, respectively). Both bDNA and LH results correlated highly with alanine aminotransferase (ALT) values (both had Prob > F values of 0.0) while PCR was not correlated with ALT (Prob > F = 0.05). In 26 evaluable patients, a model based on a generalized Knodell score was used to predict response to IFN therapy, as defined by normalization of ALT values during therapy. This model discriminated well between non-responders and responders. The bDNA results correlated well with the generalized Knodell score, while the LH results did not (Prob > F = 0.04 and 0.19 for the bDNA and LH assays, respectively). In conclusion, the bDNA assay appears to be useful for quantification of HBV DNA levels in HBeAg-negative chronic hepatitis as it correlates with biochemical and histological indications of disease severity as well as with response to IFN therapy.

Quantitation of HBV-DNA is the most precise test for measuring viral replication. A commercial liquid phase hybridation test (Abbott) is now commonly used for diagnosis and monitoring of chronic hepatitis B. Interpretation of weak positive results obtained with this test are often difficult. Fifty-four sera with a concentration lower than 12 pg/ml with the Abbott HBV-DNA assay were tested with another commercial hybridation assay (Digene-Murex) and with an in-house PCR test. PCR is positive in 24 sera among the 35 HBs antigen positive sera, but is always negative in HBs Antigene negative sera. All the HBe Antigen positive sera were positive with the PCR test. A positive result was obtained with the Digene test in only 14 sera, 13 of them were confirmed by PCR. Ten sera among the remaining 11 PCR positive sera had a low HBV-DNA concentration but under the Digene cut-off level (10 pg/ml). The sensitivity could be greatly enforced with a lower cut-off level without any lack of specificity. The PCR test remains very helpful for sera with low concentration of HBV-DNA.

To improve the sensitivity of hepatitis C virus RNA (HCV RNA) detection in serum by 'nested' polymerase chain reaction (PCR), primers belonging to 5' non-coding (5'NC) regions were used to compare the classical phenol/chloroform technique by using the proteinase K and silica gel technique with guanidinium thiocyanate. The silica gel techniques was found to be more efficient and sensitive for the extraction and purification of viral RNA from serum samples. The silica gel technique also avoids contact with hazardous volatile chemicals like phenol and chloroform and provides a better protection for viral RNA. Furthermore, the RNA detection sensitivity was greatly improved by modifying the buffer for reverse transcription and PCR. Using silica gel extraction, and the modified buffer, viral RNA was detected in 699 sera from anti-HCV second generation ELISA positive patients. These sera were distributed in second generation RIBA confirmed, indeterminate and non confirmed groups with PCR positive rates of 71.4%, 45.7% and 15.8%, respectively. Two out of 227 ELISA negative patients showed the presence of HCV RNA in serum. An association between the presence of antibodies against a determined viral peptide and viremia was not detected.

Objectives: Seroprophylaxy against hepatitis A has been limited to subjects visiting endemic areas or in contact with acutely infected patients due to the short half-life and high cost of the non-specific immunoglobulins. Inactivated vaccines might offer a better solution, but the real cost factors remain unknown. We undertook this study to determine the cost of acute hepatitis A., Methods: Serum samples from all the non-hospital medical laboratories in the Lyon (France) area were prospectively screened to identify 100 consecutive patients with acute hepatitis (positive for hepatitis A IgM antibodies). A questionnaire was addressed to these 100 subjects 1 year later requesting information on health care expenditures and work loss related to their episode of acute hepatitis A. Imprecise answers were completed by telephone interview., Results: Answers were obtained from 92 subjects. Confirmation was obtained that the 8 other subjects had not died from their episode of acute hepatitis. Only two patients were hospitalized (for 2 and 4 days). Total mean cost (including medical expenditures and work loss was 11,789 French francs (range 1,524-62,117). Medical consultations and prescriptions comprised 20.0% of the total cost. The real mean cost of work loss (8,423 F) was greater than the cost calculated by the national health service (6,500 F)., Conclusions: This evaluation of the real cost of hepatitis A in non-hospitalized patients should contribute to the definition of an adapted policy for the prevention of hepatitis A.

This paper reports the results of a survey conducted on smoking among 1490 eleven years old schoolchildren. They answered to an individual questionnaire before and after they had been informed on smoking by school doctors. In this age group, 26.2% of the children have already had some contact with cigarette, and 0.5% smoke regularly. The household smoking habits have an important influence on the attitude of children toward smoking. Knowledges on smoking dangers have been evaluated before and after information. This survey shows the importance of smoking prevention, especially among young children.

Changes in HIV replication during progression of HIV infection were assessed by estimation of the number of peripheral blood mononuclear cells (PBMC) harboring HIV proviral sequences. Samples from 23 patients at different stages of HIV infection were analyzed by quantitative polymerase chain reaction (PCR) using GAG primers. HIV titers in PBMC were also determined by serial dilutions of cells in coculture with phytohemagglutinin-activated normal PBMC. A positive correlation was observed between the number of HIV DNA copies and the HIV titer in PBMC. The PCR test was more sensitive than the coculture technique. The number of HIV copies detected by PCR ranged from 50 to 10,500 per 10(6) PBMC: assuming one copy per cell this implies a frequency of proviral HIV-containing cells of one per 100 to one per 20,000 mononuclear cells. The mean number of HIV DNA sequences in PBMC was significantly lower in asymptomatic patients than in AIDS patients and patients with AIDS-related complex (ARC). In patients who progressed from asymptomatic infection to AIDS, the number of HIV DNA copies in PBMC rose, indicating an increase of HIV replication. These results show that the number of infected PBMC increases during clinical progression. However, some asymptomatic patients had a higher number of HIV DNA copies in their PBMCs suggesting that increased HIV replication precedes the appearance of clinical symptoms.

Two mobile units for air filtration with recycling designed to reduce the aerobiocontamination of hospital rooms were tested. Their efficacity was evaluated by measurement of particle retention and by quantitative determination of the reduction of aerobiocontamination in rooms occupied by patients. Results show a significant improvement in the aerobiocontamination of test rooms as compared to control rooms. It is thus reasonable to suppose that these systems may provide a practical solution to the problems posed by hospitalization in ordinary rooms of patients subjected to aplasive chemotherapy. A randomized clinical trial is in progress.

Two hundred and twenty-eight blood samples from 154 HIV-seropositive subjects were investigated for the presence of HIV in peripheral blood mononuclear cells (PBMC) and plasma (without prior ultracentrifugation or filtration), using normal PBMC as the target for replication. HIV was recovered from PBMC and plasma in 80.5 and 19.5% of the patients, respectively. Plasma viraemia was significantly associated with clinical manifestations of HIV infection, indicating that HIV replication increased as disease progressed. This was confirmed by the statistically significant correlations between plasma viraemia and low CD4 cell counts (P less than 0.01) and low anti-p24 antibody titers (P less than 0.01). On patient follow-up, detection of HIV in plasma was transient. p24 antigenaemia was only detected in 13.6% of cases and was also associated with advanced clinical stages of the disease. When HIV RNA detection by polymerase chain reaction (PCR) was compared with plasma viraemia, HIV RNA was detected in plasma in all symptomatic cases and in 53.8% (seven out of 13) of asymptomatic patients [Centers for Disease Control (CDC) stages II and III], confirming that PCR was far more sensitive than plasma culture. These results indicate that cell-free virus production is associated with the clinical stage of HIV infection and may serve as a marker for disease progression. Detection of HIV RNA by PCR appears to be the most sensitive method to detect viraemia.

An early and accurate diagnosis of HIV infection is needed in the offspring of seropositive mothers. To this end, we have used two techniques for the direct detection of HIV in 12 newborns tested within 2 weeks after birth and 12 children. HIV isolation was carried out in lymphocyte cocultures and compared with detection of DNA and RNA sequences by molecular amplification using the polymerase chain reaction (PCR). In lymphocyte cocultures, HIV was isolated in 8 of 24 cases (33%), including 3 newborns, 3 symptomatic children, and 2 asymptomatic ones. HIV DNA was detected by PCR in twice as many cases, i.e., in 16/24 cases (66%), including 7/12 newborns, 4/4 symptomatic children, and 5/8 asymptomatic ones, 2 of whom became seronegative, HIV RNA was detected in 10 of 16 cases (60%) with detectable HIV DNA, including all of the cases who had a positive HIV isolation. Only children with clinical or biological signs of HIV infection were positive for HIV RNA. Furthermore, signs of HIV infection appeared within 6 months in three of the four newborns who were positive for HIV RNA at birth. These results indicate that HIV DNA detection by PCR is far more sensitive than HIV isolation in culture for the early diagnosis of HIV infection in offspring of seropositive mothers. HIV RNA detection appears to be a useful prognostic marker since it does correlate with disease progression and may serve as a clue for HIV replication in vivo.

After classifying the nosocomial urinary tract infections, between the hospital acquired infections, this paper review different prevention means proposed to limit their incidence. These consist essentially of recommendations concerning the vesical catheterism. Thus are reviewed catheter indications, the choice of the catheter, its putting conditions, the creation and conservation of a closed system. Besides, patient informations and bacteriological survey of the catheterized patients are also discussed.

Hospital or nosocomial infection, or infection acquired in hospitals, is a health problem in all hospital departments and particularly in the maternity department. We report on a prospective survey of surveillance of hospital-acquired infections both from the mother and the baby's point of view after delivery vaginally or with caesarean carried out at the obstetrical clinic of the Edouard Herriot Hospital in Lyon (France) over three successive years with a series of 9,204 deliveries. The incidence of infection in women who were delivered without caesarean section was 1.37% when urinary tract infections had been excluded but 13% in women who had caesarean sections. Endometritis, skin infections and urinary tract infections were the leading causes. As far as the newborn were concerned, hospital infection ran at about 2.60% and this in the main was due to staphylococcal pustules in the skin. These figures are still too high and prevention should be based on more information given and more care taken by the whole staff of such a hospital.

Some recommendations for the prevention of nosocomial urinary tract infections are irrefutable, thoroughly admitted. They concern before all the aseptic conditions of setting and care of catheter, to avoid the bacterial penetration in the closed system. They concern the merits of catheter indications, the diminution of catheter conservation period, the applying of a strict asepsis during setting and manipulation of the catheter, the use of a sterile and closed drainage system with a continuous flow. Several improvements concerning the catheter and the drainage system have been proposed. Some of these recommendations have been proved serious and can be applied. On the contrary, the efficiency of others remain to be proved by control assays. Finally, measures with uncertain or unproved values should not be considered.

We conducted in 1983 an hepatitis surveillance programme in collaboration with 93% of the medical analysis laboratories in Lyons' urban area (1,100,000 inhabitants) and diagnosed 1,002 cases of acute hepatitis (incidence : 90.5 cases/10(5) inhabitants; HAV : 50.4 cases/10(5) inhabitants; HBV : 12 cases/10(5) inhabitants; non A non B : 24 cases/10(5) inhabitants; drugs : 4.2 cases/10(5) inhabitants). HBs Ag was undetectable in 12% of acute hepatitis B. Hepatitis A accounted for 91% of children's cases and 48% of cases among adults between 20 and 40 years old. Epidemics were observed among children after summer holidays related to the return of migrants from trips to their native country. For acute hepatitis B and non A non B, classical epidemiological data were observed, but the prevalence of drug addicts and homosexuals was low.