[Sleeping Sickness: A Cause of False Positive HIV Rapid Diagnostic Tests]. / Maladie du Sommeil: Une Cause de Faux Positifs des Tests de Diagnostic Rapide du VIH.

Approaching the mechanisms related to false positives HIV rapid diagnostic tests (RDT) in patients with sleeping sickness may help to improve the accuracy of screening for HIV infection in areas endemic for Human African trypanosomiasis (HAT).We report on a patient from Congo who was managed like an AIDS-associated meningoencephalitis, based on a false positive HIV RDT at admission, and eventually received a diagnosis of sleeping sickness. A further retrospective cohort study performed in patients with HAT shows that most of positive HIV RDT obtained prior to treatment for sleeping sickness are false positives. We found that half of them were cleared at the end of treatment course, suggesting an early clearance of some antibodies involved in cross-reactivity.A substantial clearance of HIV RDT false positives occurs during therapy for HAT. In areas where Elisa HIV tests are not readily available, repeating the HIV RDT at the end of therapy may help to identify roughly half of false positives.

Characteristics of human metapneumovirus infection in adults hospitalized for community-acquired influenza-like illness in France, 2012-2018: a retrospective observational study.

OBJECTIVES: To describe the prevalence, clinical features and complications of human metapneumovirus (hMPV) infections in a population of adults hospitalized with influenza-like illness (ILI). METHODS: This was a retrospective, observational, multicenter cohort study using prospectively collected data from adult patients hospitalized during influenza virus circulation, for at least 24 h, for community-acquired ILI (with symptom onset <7 days). Data were collected from five French teaching hospitals over six consecutive winters (2012-2018). Respiratory viruses were identified by multiplex reverse transcription polymerase chain reaction (RT-PCR) on nasopharyngeal specimens. hMPV + patients were compared with hMPV- patients, influenza+ and respiratory syncytial virus (RSV)+ patients using multivariate logistic regressions. Primary outcome was the prevalence of hMPV in patients hospitalized for ILI. RESULTS: Among the 3148 patients included (1449 (46%) women, 1988 (63%) aged 65 and over; 2508 (80%) with chronic disease), at least one respiratory virus was detected in 1604 (51%, 95% confidence interval (CI) 49-53), including 100 cases of hMPV (100/3148, 3% 95% CI 3-4), of which 10 (10%) were viral co-infection. In the hMPV + patients, mean length of stay was 7 days, 62% (56/90) developed a complication, 21% (14/68) were admitted to intensive care unit and 4% (4/90) died during hospitalization. In comparison with influenza + patients, hMPV + patients were more frequently >65 years old (adjusted odds ratio (aOR) = 3.3, 95% CI 1.9-6.3) and presented more acute heart failure during hospitalization (aOR = 1.8, 95% CI 1.0-2.9). Compared with RSV + patients, hMPV + patients had less cancer (aOR = 0.4, 95% CI 0.2-0.9) and were less likely to smoke (aOR = 0.5, 95% CI 0.2-0.9) but had similar outcomes, especially high rates of respiratory and cardiovascular complications. CONCLUSIONS: Adult hMPV infections mainly affect the elderly and patients with chronic conditions and are responsible for frequent cardiac and pulmonary complications similar to those of RSV infections. At-risk populations would benefit from the development of antivirals and vaccines targeting hMPV.

Clinical characteristics and outcome of respiratory syncytial virus infection among adults hospitalized with influenza-like illness in France.

OBJECTIVES: The aim of this study was to analyse characteristics and outcome of respiratory syncytial virus (RSV) infection in adults hospitalized with influenza-like illness (ILI). METHODS: Patients hospitalized with ILI were included in this prospective, multicentre study carried out in six French hospitals during three consecutive influenza seasons (2012-2015). RSV and other respiratory viruses were detected by multiplex PCR in nasopharyngeal swabs. Risk factors for RSV infection were identified by backward stepwise logistic regression analysis. RESULTS: A total of 1452 patients hospitalized with ILI were included, of whom 59% (861/1452) were >65 years and 83% (1211/1452) had underlying chronic illnesses. RSV was detected in 4% (59/1452), and influenza virus in 39% (566/1452). Risk factors for RSV infection were cancer (adjusted OR 2.1, 95% CI 1.1-4.1, p 0.04), and immunosuppressive treatment (adjusted OR 2.0, 95% CI 1.1-3.8, p 0.03). Patients with RSV had a median length of stay of 9 days (6-25), and 57% of them (30/53) had complications, including pneumonia (23/53, 44%) and respiratory failure (15/53, 28%). Fifteen per cent (8/53) were admitted to an intensive care unit, and the in-hospital mortality rate was 8% (4/53). Pneumonia was more likely to occur in patients with RSV than in patients with RSV-negative ILI (44% (23/53) versus 26% (362/1393), p 0.006) or with influenza virus infection (44% versus 28% (157/560), p 0.02). CONCLUSION: RSV is an infrequent cause of ILI during periods of influenza virus circulation but can cause severe complications in hospitalized adults. Risk factors for RSV detection in adults hospitalized with ILI include cancer and immunosuppressive treatment. Specific immunization and antiviral therapy might benefit patients at risk.

[Evaluation of accuracy of three assays for human papillomavirus detection and typing: Hybrid Capture 2, HPV Consensus kit and Amplicor HPV]. / Evaluation des performances de trois techniques de détection et de typage des papillomavirus humains: Hybrid Capture 2, HPV Consensus kit et Amplicor HPV.

BACKGROUND: Detection of high-risk human papillomavirus has proved its usefulness in complement of abnormal cervical scrape result. The Hybrid Capture 2 (HC2, Digene) test has proven its efficiency. We have compared this test with HPV Consensus kit (HPVC, Argène) and Amplicor HPV test (AHPV, RocheDiagnostics) on a panel of 88 samples with low HC2 ratios or discordant results between HC2 and cervical scrape. MATERIAL AND METHODS: Cervical samples were tested in parallel by the three methods using a nested amplification of L1 region as reference. RESULTS: Eighty-six samples were suitable for analysis. Results of HC2 and AHPV tests were closely related. The use of a "generic" probe in the HPVC test was responsible for undetermined results, which were not clinically relevant. CONCLUSION: Despite the low viral load of the samples chosen, the hybridization (HC2) and PCR (AHPV or HPVC) methods gave comparable results, with false positive and false negative results for all tests, but a 75% concordance and a high sensibility to detect HPV infection. However, a complementary study on a larger population with ASCUS diagnosis and biopsy under colposcopy would be necessary to valid these assays for a clinical indication.

[Herpesviruses serologic survey of corneal allograft recipients]. / Surveillance sérologique pour les Herpesvirus chez les receveurs de cornée.

AIMS: Herpesviruses are ubiquitous viruses, providing circulating antibodies in a wide range of patients. Donor-to-host transmission of Herpes simplex virus via corneal graft has been proven, leading to primary graft failure. However, the serological survey of the corneal recipient for Herpesviruses has not yet been investigated. METHODS: Circulating antibodies to HSV, VZV, CMV, and EBV were tested in 117 corneal recipients prior to surgery as well as 8 days and 3 months following surgery. Twenty-two patients had a history of corneal herpes. All patients were treated with local steroids, and no patient received systemic immunosuppressive therapy. RESULTS: No seroconversion was encountered, in particular, no CMV--patient was found CMV+ after grafting. The mean concentration of antibodies significantly decreased after grafting in a few patients. A serological profile of EBV reactivation was detected after surgery in four patients at day 8 and three more patients at 3 months. CONCLUSIONS: This study shows no significant seroconversion after grafting. However, it shows a postoperative decrease in antibody levels as well as a serological profile of EBV reactivation, possibly related to local steroids or graft immune processes.

[Hybrid Capture 2 vs HPV Consensus kit: comparison of 2 assays for human papillomavirus detection and typing]. / Comparaison de deux techniques de détection et de typage des papillomavirus humains: Hybrid Capture 2 et HPV Consensus kit.

BACKGROUND: Many laboratories use the DNA Hybrid Capture 2 HPV-high risk assay (Digene) to detect and type oncogenic HPV. The aim of this work was to compare this assay with a new HPV genotyping assay: HPV Consensus kit (Argène). Actually, this assay is not commercially available. MATERIALS AND METHODS: Ninety-four cervical samples were tested with both the routine assay Hybrid Capture 2 and the HPV Consensus kit. Discordant results were analysed by amplification with a nested PCR and sequencing of amplified products. RESULTS: Only 81 results could be analysed concerning the oncogenic risk. The overall concordance was 92,6%. But we find 13 "generic" results with the HPV Consensus kit, the generic probe including high risk and low risk genotypes. CONCLUSION: HPV Consensus kit results showed a better detection sensitivity for this assay than Hybrid Capture 2 assay. Nevertheless, "generic" results give no information about the oncogenic risk of the HPV detected in a sample.

Human tumor suppressor p53 and DNA viruses.

Human tumor suppressor protein p53 plays a major role in the cell cycle, orchestrating a number of important genes involved in cell-cycle control and apoptosis, and seems to be one of the most important molecules protecting cells from malignant transformation. Mutations in the p53 gene are observed in about 50% of primary tumors, inducing defective p53 protein no longer capable of binding DNA and of activating transcription. Certain DNA viruses are thought to act in a similar way and may also contribute to the progression of invasive cancer in infected tissue. One of the most effective strategies employed by these viruses is the inhibition of p53 protein by interaction with viral oncoproteins, implying a direct but also an indirect role of these viruses in the impairment of p53 structure and function. This article provides a summary of current knowledge concerning p53 tumor suppressor protein and reviews the different mechanisms adopted by different DNA viruses in undermining p53 function.

Detection of ganciclovir resistance after valacyclovir-prophylaxis in renal transplant recipients with active cytomegalovirus infection.

Whether valaciclovir (VCV) prophylaxis could be responsible for ganciclovir (GCV)-resistance of Human cytomegalovirus (HCMV) in transplantation has never been documented. A multicentric retrospective pilot study was undertaken to detect GCV-resistance through mutations within the UL97 gene in renal transplant recipients who experienced active HCMV infection and received valacyclovir prophylaxis. Twenty-three patients who experienced HCMV antigenaemia or DNAemia during or at the end of prophylaxis were included. UL97 genotyping was carried out on peripheral blood samples, using a nested in-house PCR, which amplified the full-length UL97 gene. One patient has a resistance-related mutation (M460I); the major risk factor for emergence of resistance in this patient was the presence of early and persistent antigenaemia. GCV-resistance during VCV-prophylaxis was rare after renal transplantation. However, special attention must be paid to patients developing early active HCMV infection under prophylaxis.

[Antiviral therapy adjustment in corneal recipients using antibody testing in the aqueous humor]. / Ajustement du traitement antiviral après greffe de cornée par mesure de la production d'anticorps dans l'humeur aqueuse.

INTRODUCTION: In corneal recipients with herpes infection, acyclovir given for 1 year postoperatively prevents viral reactivation and improves graft outcome. The indication for prophylactic antiviral therapy relies on the preoperative diagnosis of herpes. However, many patients present with corneal scars featuring sequelae of herpes without a proven history of herpes. Here we report the results of a prospective study of anti-herpes simplex virus (anti-HSV) and varicella zoster virus (VZV) antibody testing in the aqueous humor at the time of corneal transplantation to refine the indication of the antiviral treatment. MATERIAL AND METHODS: The study involved 33 keratitis corneal graft recipients, 21 of whom had documented herpes keratitis. A control group was made with 11 cataract patients. An anterior chamber puncture was performed just before surgery. The micro-ELISA test was done on both aqueous humor and serum, and local anti-HSV or VZV antibody synthesis was acknowledged if the ratio of antibody concentrations was above 4. RESULTS: Local antibody synthesis to HSV was detected in 22 cases, to VZV in 9 cases, to both HSV and VZV in 6 cases, and no synthesis in 8 cases. The sensitivity of the test was 65% in patients with a documented history of herpes (14 cases out of 21). Among non-herpetic patients, the test was positive in 9 patients, who thus benefited from postoperative antiviral therapy. No viral reactivation was encountered after a minimum follow-up of 1 year. CONCLUSIONS: Antibody testing in the aqueous humor at the time of keratoplasty is a convenient, inexpensive diagnostic tool in corneal recipients. It provides useful information before prescribing a long and expensive postoperative antiviral therapy.

Drug-induced hypersensitivity syndrome associated with Epstein-Barr virus infection.

Association of drug-induced hypersensitivity syndrome with viral infection is debated. Human herpesvirus 6 (HHV-6) reactivation has been the most frequently reported infection associated with this syndrome. However, a case of cytomegalovirus (CMV) infection was recently described associated with anticonvulsant-induced hypersensitivity syndrome. We report a case of severe allopurinol-induced hypersensitivity syndrome with pancreatitis associated with Epstein-Barr virus (EBV) infection. Active EBV infection was demonstrated in two consecutive serum samples by the presence of anti-EBV early antigen (EA) IgM antibodies and an increase in anti-EBV EA IgG antibodies, whereas no anti-EBV nuclear antigen IgG antibodies were detected. EBV DNA was detected by polymerase chain reaction (PCR) in peripheral blood mononuclear cells. Reactivation of HHV-6 was suggested only by the presence of anti-HHV-6 IgM antibodies, but HHV-6 DNA was not detected by PCR in the serum. Other viral investigations showed previous infection (CMV, rubella, measles, parvovirus B19), immunization after vaccination (hepatitis B virus), or absence of previous infection (hepatitis C virus, human immunodeficiency virus). We suggest that EBV infection may participate in some cases, as do the other herpesviruses HHV-6 or CMV, in the development of drug-induced hypersensitivity syndrome.

Real-time PCR for quantification of human herpesvirus 6 DNA from lymph nodes and saliva.

A real-time quantitative PCR assay has been developed to measure human herpesvirus 6 (HHV-6) DNA in biological specimens. The assay sensitivity was 10 copies of DNA per well, with a linear dynamic range of 10 to 10(7) copies of HHV-6 DNA. Intra- and interassay variations were, respectively, 0.88 and 0.8% for samples containing 10(2) DNA copies, 0.99 and 0.96% for samples containing 10(4) copies, and 0.76 and 0.9% for samples containing 10(6) copies. Among 34 saliva samples from healthy subjects, 26 were found to contain HHV-6 DNA (76.5%; median, 23,870 copies/ml), and following a single freeze-thaw cycle, 25 of the same samples were found to be positive for HHV-6 DNA, although at a statistically significantly lower concentration (median, 3,497 copies/ml). The assay enabled detection of HHV-6 DNA in lymph node biopsies from patients with Hodgkin's disease (HD) (13 of 37 patients [35.1%]), B-cell neoplasms (8 of 36 patients [22.2%]), and T- or NK-cell neoplasms (3 of 13 patients [23.1%]), with concentrations ranging from 100 to 864,640 HHV-6 copies per microg of DNA (HHV-6B being found in every case except two). All HD patients infected with HHV-6 presented clinically with the nodular sclerosis subtype of HD. The real-time quantitative PCR assay developed here was simple to perform and was sensitive over a wide range of HHV-6 concentrations. It therefore appears to be of potential value in clinical investigation or diagnosis of HHV-6 infection.

Prevalence and characteristics of Sjögren's syndrome or Sicca syndrome in chronic hepatitis C virus infection: a prospective study.

OBJECTIVE: To describe the prevalence and clinical and laboratory characteristics of sicca syndrome and Sjögren's syndrome (SS) in chronic hepatitis C virus (HCV) infection. METHODS: Forty-five consecutive HCV infected patients referred for liver biopsy were enrolled in a prospective study. Subjective and objective criteria of xerophthalmia or xerostomia were systematically investigated and the patients classified according to 3 sets of criteria (European, Manthorpe, and Fox criteria) for the diagnosis of SS. RESULTS: Sicca syndrome was present in 28 (62%) patients; all had oral dryness and 14 had both oral and ocular dryness. Twenty-four (53%) patients had SS by the European criteria, 25 (56%) by Manthorpe criteria, and 4 (8%) by Fox criteria. Salivary gland biopsy was positive for SS (grade III or IV by Chishom classification) in 21 samples (47%); 9 samples (21%) were classified grade 0, and 15 (32%) grade I or II. No patient had anti-SSA or anti-SSB antibodies. The presence of SS or sicca syndrome was associated with older age and liver disease activity according to the METAVIR scoring system, but not with the presence of other extrahepatic manifestations or with HCV genotype. A high METAVIR activity score was only statistically associated with primary SS. CONCLUSION: HCV infection appears to account for a subgroup of patients with sicca syndrome in which half the cases meet the definition for SS according to European and Manthorpe criteria. This subgroup is characterized by the constant finding of xerostomia, the absence of classical systemic manifestations observed in primary SS, and the absence of anti-SSA or anti-SSB antibodies. Such characteristics delineate a distinctive, virus associated entity that differs from primary SS.

Evaluation of BC-3 and BCP-1 cell lines in immunofluorescence assay to detect HHV-8 antibodies in Kaposi's sarcoma, multiple myeloma and other groups.

BACKGROUND: We describe a comparative study of an immunofluorescence assay using inducible BC-3 and BCP-1 cell lines as sources of HHV-8 antigens. STUDY DESIGN: Detection of both antibodies to proteins expressed in lytic cycle and during latency in sera from HIV-infected patients with KS, HIV-positive patients without KS, normal blood donors, HIV-negative pregnant women and HIV-negative patients with multiple myeloma. Where possible, detection of antibody was associated with nested PCR detection of HHV-8 in peripheral mononuclear cell (PBMC) samples collected from AIDS-KS patients. RESULTS: Immunofluorescence was more intense with the BC-3 cell line than with BCP-1, thus facilitating examination under the microscope. HHV-8 antibodies were detected among 82.75% of AIDS-KS patients, in 27.3% of HIV-infected homosexual men, 2% of blood donors and in 2% of pregnant women. No HHV-8 antibodies were detected in serum samples from HIV-negative patients presenting multiple myeloma. HHV-8 DNA sequences were detected and confirmed by southern blot hybridization in five out of 17 (29.4%) PBMC samples from AIDS-KS patients. Titre of antibodies to proteins expressed in lytic cycle was much higher than the titre of antibodies to proteins expressed during latency. CONCLUSIONS: Both immunofluorescence assays were found useful and HHV-8 seroprevalence rates reported in previous studies were confirmed. In addition, results obtained using these assays tend to provide evidence for a lack of epidemiological association between HHV-8 infection and development of multiple myeloma.

[Human herpesviruses 6 and 7 (HHV-6 and HHV-7)]. / Herpesvirus humains 6 et 7 (HHV-6 et HHV-7).

Human herpesviruses 6 (HHV-6) and 7 (HHV-7) are world wide T lymphotropic viruses recently discovered. Their transmission is essentially by salivary route. Primary infections which occurred early in infancy, respectively during the first year of life for HHV-6 and in the second or third year for HHV-7, are followed by latency for life. HHV-7 is not actually associated with a disease. HHV-6 primary infection is often asymptomatic, if not it can induce exanthem subitum. HHV-6 reactivation can be symptomatic in immunodeficient subjects. The role of HHV-6 in the arising of lymphoproliferative or auto-immune diseases, discussed for a long time, is still to elucidate. HHV-6 infection is diagnosed by serodiagnosis in case of primary infection, but in the great number of cases, it would be realized by polymerases chain reaction.

[Virus and gastrointestinal infections]. / Virus et infections gastro-intestinales.

Numerous viruses found in the gut are not associated with primary infection or disease of the gastrointestinal tract. Other groups or viruses are not classically associated with infection of the gut but can infect the gastrointestinal tract in immunocompromised individuals (herpes simplex virus, cytomegalovirus, papillomavirus ....). The viruses associated with gastroenteritis represent a large number of taxonomic group. Because these viruses have in general been difficult to cultivate, most members of this group were firstly detected by electron microscopic examination (adenovirus, astrovirus, calicivirus, coronavirus, rotavirus ....). The most widely used diagnostic techniques for adenovirus (40/41), rotavirus and astrovirus detection in faecal samples include immunoassays such as Elisa and latex agglutination. Nucleic acid hybridization techniques have generally not proven to be substantially sensitive and the more sensitive techniques recently developed use the polymerase chain reaction (adenovirus) or the reverse transcription/polymerase chain reaction (astrovirus, calicivirus, coronavirus, rotavirus). Special efforts have been made in the search for efficient procedures to extract viral nucleic acids, and to establish the optimal conditions for the amplification and identification of PCR products but the candidate viruses were very different, consensus procedures were not determined, and amplification kits were not actually commercialized.

Evaluation of rapid culture centrifugation method for adenovirus detection in stools.

Routine laboratory testing for adenovirus (Adv) requires a procedure that is rapid and reliable, especially for samples from children and immunosuppressed patients, when diarrhea may signal the onset of severe gastrointestinal disorders. An improved culture technique for Adv isolation, using centrifugation step of 24-well plates and needing only 48 h incubation, was evaluated for 382 stool samples. This technique was compared with conventional tube cell culture and a commercial enzyme-linked immunosorbent assay (ELISA) kit. Adv was isolated in 36 samples (9.4%) by rapid culture, in 32 (8.4%) by conventional culture, and in 42 samples (11%) using genus-specific ELISA. A total of 30 isolates were found to be Adv positive in both rapid and conventional cultures, and half of the Adv-positive rapid culture isolates were identified as serotypes 40/41 using a type-specific ELISA. The improved culture method considerably reduces incubation time and also offers a slightly enhanced sensitivity to Adv serotypes. Combined with appropriate cell lines adapted to the isolation of enteric adenoviruses, it therefore constitutes a valuable laboratory test particularly useful in the diagnosis of gastroenteritis.

[Adenoviruses from stool samples in hospital units. Comparison with main pathogens in gastroenteritis (rotavirus, Campylobacter, Salmonella)]. / Les adénovirus dans les prélèvements de selles en milieu hospitalier. Comparison avec les principaux agents des gastroentérites (rotavirus, Campylobacter, Salmonella).

During a six-years period (1988-1993), a total of 14,644 stool samples from in-patients of Limoges University Hospital were examined for the presence of principal enteric pathogens, such as adenovirus, rotavirus, Campylobacter, Salmonella, Shigella and others. Stools were processed for identification of bacteria by standard methods and viruses were detected in fecal specimens using antigen detection methods: ELISA (Enzyme Linked Immunosorbent Assay) and latex agglutination test. The decreasing rates of presence of enteric agents were respectively 6% for rotavirus, 3.2% for Salmonella, 2% for adenovirus, 1.6% for Campylobacter and 0.2% for Shigella, but according to the lack of sensibility of latex agglutination test, adenovirus prevalence was probably underestimated. Concerning the distribution of enteric pathogens throughout the year, our data demonstrate that rotavirus were rather shed during the months from January to April, adenovirus between April and August, Campylobacter during summer and Salmonella from July to October. The two thirds of Campylobacter and rotavirus infections and the half of adenovirus and Salmonella infections were identified during the ten first years of life. The highest prevalence occurs before 5 years old, during the 2nd year of life for adenovirus (4.4%) and rotavirus (22.3%) and during the 3rd year of life for Campylobacter (6.84%) and Salmonella (8.6%).