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1.
Clin Infect Dis ; 72(9): 1631-1634, 2021 05 04.
Article in English | MEDLINE | ID: mdl-32614433

ABSTRACT

Prolonged measles virus detection in maternal saliva and blood was evidenced in 6 pregnant women. Maternal-fetal transmission was evidenced in 2 of 4 infants who were asymptomatic at birth, 21-24 weeks after maternal infection. Whereas peripartum congenital measles is severe, asymptomatic measles virus vertical transmission can occur earlier in pregnancy.


Subject(s)
Measles , Pregnancy Complications, Infectious , Female , Humans , Infant , Infant, Newborn , Infectious Disease Transmission, Vertical , Measles virus , Parturition , Pregnancy
2.
Viruses ; 8(8)2016 08 20.
Article in English | MEDLINE | ID: mdl-27556477

ABSTRACT

The objective of this study is primarily to compare the performance of the VIDAS(®) Measles immunoglobulin (Ig)G assay to that of two other serological assays using an immunoassay technique, Enzygnost(®) Anti-measles Virus/IgG (Siemens) and Measles IgG CAPTURE EIA(®) (Microimmune). The sensitivity and the agreement of the VIDAS(®) Measles IgG assay compared to the Enzygnost(®) Anti-measles Virus/IgG assay and the Measles IgG CAPTURE EIA(®) assay are 100%, 97.2% and 99.0%, 98.4%, respectively. The very low number of negative sera for IgG antibodies does not allow calculation of specificity. As a secondary objective, we have evaluated the ability of the VIDAS(®) Measles IgG assay to measure anti-measles virus IgG antibody avidity with the help of the VIDAS(®) CMV IgG Avidity reagent, using 76 sera from subjects with measles and 238 other sera. Different groups of populations were analyzed. In the primary infection measles group, the mean IgG avidity index was 0.16 (range of 0.07 to 0.93) compared to 0.79 (range of 0.25 to 1) in the serum group positive for IgG antibodies and negative for IgM. These data allow to define a weak anti-measles virus IgG antibody avidity as an avidity index (AI) < 0.3 and a strong avidity as an AI > 0.6. The VIDAS(®) Measles IgG assay has a performance equivalent to that of other available products. Its use, individual and quick, is well adapted to testing for anti-measles immunity in exposed subjects.


Subject(s)
Antibodies, Viral/blood , Antibody Affinity , Immunoassay/methods , Immunoglobulin G/blood , Measles virus/immunology , Measles/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Sensitivity and Specificity , Young Adult
3.
Emerg Infect Dis ; 19(3): 357-64, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23618523

ABSTRACT

Although few measles cases were reported in France during 2006 and 2007, suggesting the country might have been close to eliminating the disease, a dramatic outbreak of >20,000 cases occurred during 2008-2011. Adolescents and young adults accounted for more than half of cases; median patient age increased from 12 to 16 years during the outbreak. The highest incidence rate was observed in children <1 year of age, reaching 135 cases/100,000 infants during the last epidemic wave. Almost 5,000 patients were hospitalized, including 1,023 for severe pneumonia and 27 for encephalitis/myelitis; 10 patients died. More than 80% of the cases during this period occurred in unvaccinated persons, reflecting heterogeneous vaccination coverage, where pockets of susceptible persons still remain. Although vaccine coverage among children improved, convincing susceptible young adults to get vaccinated remains a critical issue if the target to eliminate the disease by 2015 is to be met.


Subject(s)
Disease Outbreaks/prevention & control , Mass Vaccination , Measles/prevention & control , Adolescent , Adult , Child , Child, Preschool , Cross Infection/epidemiology , Cross Infection/prevention & control , Cross Infection/virology , Female , France/epidemiology , Genotype , Humans , Infant , Male , Measles/epidemiology , Measles/virology , Measles Vaccine , Morbillivirus/genetics , Morbillivirus/immunology , Young Adult
4.
J Med Virol ; 85(4): 723-30, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23364811

ABSTRACT

During the 2011 measles outbreak in Paris (France), patients with clinical suspicion of measles were tested for virological confirmation of measles virus (MV) infection. To assess the practical value of molecular diagnosis in an epidemic setting, 171 oral fluid samples and 235 serum samples collected from 270 patients were tested prospectively for MV-RNA using a novel one-step real-time RT-PCR assay including an internal control. Serum samples were also tested for MV-specific IgG and IgM antibodies. MV infection was confirmed by detection of MV-RNA and/or MV-IgM for 229 of the 270 patients. The results for the 102 cases with both serum and oral fluid samples available were used to compare the techniques. The detection rate of MV-RNA by RT-PCR was 98% (100/102) for oral fluid and 95% (97/102) for serum samples. The detection rate of MV-IgM was 85% (87/102). Negative MV-IgM results were observed mostly for serum samples collected early after the onset of the rash. A MV-RNA standard of known concentration obtained by in vitro transcription was used to quantify MV-RNA in samples. MV-RNA copy numbers were significantly higher in oral fluid than in serum samples, but did not correlate with time of sampling (within 1 week after the onset of the rash), patient age, or vaccination status. During the early stage of infection, the MV-RNA viral load in serum was lower in patients positive than in those negative for MV-IgG. In conclusion, the one-step real-time RT-PCR assay is a simple and sensitive tool suitable for MV diagnosis within hours.


Subject(s)
Epidemics , Measles virus/isolation & purification , Measles/diagnosis , Measles/virology , Mouth/virology , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Adolescent , Adult , Aged , Antibodies, Viral/blood , Child , Child, Preschool , Female , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Infant , Male , Measles/epidemiology , Middle Aged , Molecular Diagnostic Techniques/methods , Paris/epidemiology , Sensitivity and Specificity , Serum/virology , Young Adult
5.
Med Sci (Paris) ; 28(11): 1003-7, 2012 Nov.
Article in French | MEDLINE | ID: mdl-23171908

ABSTRACT

From January 2008 to May 2012, over 22,000 cases of measles were reported in France. The highest incidence rate was observed in children below one year of age. Over 50% of cases were reported in young adults. Almost 5,000 patients were hospitalised including 1,023 with severe pneumonia, 27 with encephalitis and/or myelitis : 10 died. This situation is linked to insufficient and heterogeneous vaccination coverage with pockets of susceptible people allowing virus circulation. Although the vaccine coverage in children has now improved for both doses, the issue of convincing young susceptible adults to catch up for measles vaccination remains critical, if the elimination target is to be met, and in order to protect the most vulnerable population unable to benefit from this vaccination (children below 1 year, immunodeficient people, pregnant women).


Subject(s)
Measles/epidemiology , Adolescent , Adult , Child , Child, Preschool , Diarrhea/etiology , Encephalitis, Viral/etiology , France/epidemiology , French Guiana/epidemiology , Geographic Mapping , Guadeloupe/epidemiology , Humans , Incidence , Infant , Infant, Newborn , Martinique/epidemiology , Measles/complications , Measles/prevention & control , Measles Vaccine , Otitis Media/etiology , Pneumonia, Viral/etiology , Reunion/epidemiology , Subacute Sclerosing Panencephalitis/etiology , Vaccination/statistics & numerical data , Young Adult
6.
J Med Virol ; 83(4): 695-701, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21328385

ABSTRACT

Rapid and specific diagnosis of influenza A/B and respiratory syncytial virus (RSV) viruses is needed for optimal management of patients with acute respiratory infections. In this study, a one-step triplex real-time RT-PCR assay was developed for rapid diagnosis of influenza A/B and RSV infections to optimize diagnosis efficiency of acute respiratory infections. Cell-culture supernatants and clinical samples were used to evaluate specificity and sensitivity of the assay. The assay was used routinely during two winter epidemics for testing respiratory specimens from 2,417 patients. The limit of detection in cell-culture supernatant was 1-10 plaque forming units/input (influenza A/B) and 2 × 10(-2) 50% tissue culture infectious dose/input (RSV). In clinical samples, the assay was as sensitive as commercial molecular assays for the detection of each influenza A/B and RSV (Flu-A/B and RSV-A/B r-gene™) individually, and far more sensitive than antigen detection. During the winter 2008-2009, the assay identified 145 RSV, 42 influenza A, and one mixed RSV-influenza A infections among 298 patients. The next winter, the assay was used in two independent hospital laboratory settings. 776 patients were tested in one hospital and 1,343 in the other, resulting in 184 and 501 RSV, 133 and 150 influenza A, and 1 and 11 mixed RSV-influenza A infections, respectively, being detected. This new user-friendly assay allows rapid (within hours), effective molecular diagnosis of single or mixed infections involving influenza A (including seasonal A H1N1 and H3N2, and A(H1N1) 2009), influenza B, and RSV(A/B). The assay is very valuable for managing patients during winter epidemics when influenza and respiratory syncytial viruses co-circulate.


Subject(s)
Influenza A virus/isolation & purification , Influenza B virus/isolation & purification , Molecular Diagnostic Techniques/methods , Respiratory Syncytial Virus, Human/isolation & purification , Respiratory Tract Infections/virology , Reverse Transcriptase Polymerase Chain Reaction/methods , Virus Diseases/diagnosis , Adult , Aged , Aged, 80 and over , Child, Preschool , Humans , Infant , Influenza A virus/genetics , Influenza B virus/genetics , Middle Aged , Respiratory Syncytial Virus, Human/genetics , Respiratory Tract Infections/diagnosis , Sensitivity and Specificity , Virology/methods , Virus Diseases/virology
7.
J Med Virol ; 83(3): 517-24, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21264874

ABSTRACT

Human metapneumovirus (hMPV) is responsible for respiratory tract disease, particularly in the young and elderly population. An epidemiological and phylogenic study was performed on children admitted to hospital with an acute lower respiratory tract infection (LRI). Data were obtained and analyzed over three consecutive winters, from 2002-2003 to 2004-2005. Each year during the winter period, from November to March, 2,415 nasal swabs were tested by a direct immunofluorescence assay (DFA) for influenza viruses A and B, respiratory syncytial virus, parainfluenza viruses, and adenoviruses. Rhinoviruses, enteroviruses, and coronaviruses OC43 and 229E were detected by RT-PCR. A RT-PCR designed for the M gene was performed on negative samples for hMPV detection and phylogenic analyses. For the three consecutive winters, hMPV represented 10%, 22.6%, and 8.8% of virus-negative samples, respectively. In most cases, clinical symptoms indicated a LRI with a final diagnosis of bronchiolitis. During the winter of 2003-2004, all viral clusters (A1, A2, B1, and B2) that circulated in France shifted progressively from the A group to the B group. This study determined the prevalence of hMPV in Normandy, its clinical impact and permitted the analysis of the molecular evolution during the successive outbreaks.


Subject(s)
Disease Outbreaks , Metapneumovirus/classification , Metapneumovirus/genetics , Paramyxoviridae Infections/epidemiology , Paramyxoviridae Infections/virology , Phylogeny , Child, Preschool , Female , Fluorescent Antibody Technique, Indirect , France/epidemiology , Humans , Infant , Male , Molecular Sequence Data , Molecular Typing , Viral Matrix Proteins/genetics
8.
J Clin Virol ; 50(1): 61-4, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20952249

ABSTRACT

BACKGROUND: Human parvovirus B19 (PVB19) infection is occasionally associated with acute myocarditis. Three cases of children with PVB19 virus-associated myocarditis occurred in a very short period and the same geographical region. OBJECTIVE: To elucidate if virological factors could be responsible for determining the course of infection, a molecular epidemiologic investigation was performed. STUDY DESIGN: The diagnosis of myocarditis was established by histology or echocardiography. In the three cases, the PVB19 DNA was detected in different samples. Eight different regions were amplified by PCR using a high fidelity Taq polymerase and sequenced on both strands. Phylogenetic analyses were performed. First, the genotypes of the PVB19 strains were determined, then the intra-patient viral variability was analysed by sequencing PVB19 detected in different specimens sampled from the same patient at the same moment. RESULTS: Nearly complete sequences of the PVB19 virus (4265nt) were obtained from different samples in the three patients. The phylogenetic analyses showed that PVB19 strains identified clustered with genotype 1a PVB19 strains referenced in GenBank. When compared to the referenced strain NC_000883, the number of substitutions (transitions and transversions) were as follows: 58 for Caen.FRA/19.09, 74 for Caen.FRA/21.09 and 60 for Caen.FRA/24.09. The strains isolated from the same patient showed 100% of similarity. CONCLUSIONS: Viral myocarditis is a frequently unrecognized cause of post-inflammatory cardiomyopathy. The detailed molecular analyses do not give rise to virological markers associated with myocarditis in these children.


Subject(s)
Myocarditis/virology , Parvoviridae Infections/virology , Parvovirus B19, Human/genetics , Acute Disease , Child, Preschool , DNA, Viral/genetics , Echocardiography , Female , Genotype , Humans , Infant , Myocarditis/physiopathology , Parvoviridae Infections/physiopathology , Polymerase Chain Reaction
9.
Virologie (Montrouge) ; 15(1): 6-22, 2011 Feb 01.
Article in French | MEDLINE | ID: mdl-36151651

ABSTRACT

After the huge decrease of measles thanks to vaccination, measles reappeared in 2008, with 604 cases reported at the Institut national de veille sanitaire (InVS), then 1,544 cases in 2009 and 2,605 cases up to 2010, June. At the same time, 86 viral strains were detected from saliva samples at the Centre national de référence de la rougeole et des Paramyxoviridae respiratoires (CNR) in 2008, 316 in 2009 and 946 up to August 2010. The reality of the outbreak was confirmed by the increase of the endemic cases: 0.0009% cases in 2008 and 0.004% in 2010, the diffusion to all parts of France, and the more specific attack of infants: 4% in 2008 and 9% in 2010, and of young adults: 17% in 2008 and 38% in 2010. Most of the cases (82%) occurred in non-vaccinated people. The number of hospitalised cases has increased as well, going from 18% in 2008 to 34% in 2010. The strain of this outbreak is a genotype D4. It appeared in 2008 then it spread in 2009 and 2010, representing 19, 75 and 99% of the strains, respectively. All the viruses in this genotype belonged to the Montreal-like cluster described in 1989: Montréal.CAN/89xD4. At the beginning of the outbreak some were closed to a variant which appeared in England in 2007 MVs/Enfield.GBR/14.07(D4), but most of them (95%) are nowidentical to a strain which caused a small focus of measles in the region Vendee at the last trimester 2008: MVs/Montaigu.FRA/43.08(D4). The salivary diagnosis of measles, which was introduced in France in 2005, in parallel to the obligation of reporting measles cases, has been proved very efficient: 75% of saliva are collected in the first four days, and viral RNA was detected in 536 (81%) out the 660 samples received at the CNR up to now in 2010; 136 (21%) saliva had IgM specific antibodies and 18% had neither RNA, nor IgM.

10.
J Med Virol ; 82(10): 1762-72, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20827775

ABSTRACT

Acute respiratory infections are a major cause of mortality and morbidity worldwide. Using multiplex PCR/RT-PCR methods for the detection of 18 respiratory viruses, the circulation of those viruses during 3 consecutive dry seasons in Cambodia was described. Among 234 patients who presented with influenza-like illness, 35.5% were positive for at least one virus. Rhinoviruses (43.4%), parainfluenza (31.3%) viruses and coronaviruses (21.7%) were the most frequently detected viruses. Influenza A virus, parainfluenza virus 4 and SARS-coronavirus were not detected during the study period. Ninety apparently healthy individuals were included as controls and 10% of these samples tested positive for one or more respiratory viruses. No significant differences were observed in frequency and in virus copy numbers for rhinovirus detection between symptomatic and asymptomatic groups. This study raises questions about the significance of the detection of some respiratory viruses, especially using highly sensitive methods, given their presence in apparently healthy individuals. The link between the presence of the virus and the origin of the illness is therefore unclear.


Subject(s)
Respiratory Tract Diseases/epidemiology , Respiratory Tract Diseases/virology , Reverse Transcriptase Polymerase Chain Reaction/methods , Virology/methods , Virus Diseases/epidemiology , Virus Diseases/virology , Viruses/classification , Adolescent , Adult , Aged , Cambodia/epidemiology , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Prevalence , Viruses/isolation & purification , Young Adult
11.
Vet Microbiol ; 145(3-4): 329-33, 2010 Oct 26.
Article in English | MEDLINE | ID: mdl-20427133

ABSTRACT

Equine herpesvirus 1 (EHV-1) is a common pathogen of the horse which may induce mild respiratory distress, abortion, neonatal death and neurological disease. A single nucleotide polymorphism in the EHV-1 DNA polymerase (ORF30 A(2254) to G(2254)) has been associated with clinical signs of Equine herpes myeloencephalopathy (EHM). The aim of this work was to analyze the ORF30 genomic region among a panel of EHV-1 DNA extract in order to estimate the prevalence of the EHV-1 neuropathogenic genotype in France. Samples coming from cases associated with EHM, horses with respiratory symptoms and aborted mares, each obtained between 2002 and 2009, were investigated. DNA was directly extracted from biological samples and allelic discrimination was performed using real-time PCR. Thirty of the 125 analysed horses (24%) presented the G(2254) genotype of ORF 30. Among them, 7/16 were provided by EHM cases, 1/24 by respiratory cases and 22/85 by abortion cases. Concerning EHM, the 7 G(2254) genotype of ORF30 were all isolated in 2009 during two outbreaks where mortality was observed. Regarding the 22 G(2254) genotype of ORF 30, 17 were identified in foetuses on which EHV-1 was detected by PCR, without any certainty of viral implication in the abortion. These findings clearly suggest that other factors need to be considered for a better understanding of the impact of DNA polymerase genotype upon EHV-1 neuropathogenic phenotype.


Subject(s)
Genetic Variation/genetics , Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/isolation & purification , Horse Diseases/virology , Nervous System Diseases/veterinary , Polymorphism, Single Nucleotide/genetics , Animals , DNA, Viral/chemistry , DNA, Viral/genetics , DNA-Directed DNA Polymerase/chemistry , DNA-Directed DNA Polymerase/genetics , France/epidemiology , Genotype , Herpesviridae Infections/epidemiology , Herpesviridae Infections/virology , Herpesvirus 1, Equid/classification , Herpesvirus 1, Equid/genetics , Horse Diseases/epidemiology , Horses , Nervous System Diseases/epidemiology , Nervous System Diseases/virology , Open Reading Frames/genetics , Polymerase Chain Reaction/veterinary , Prevalence
12.
J Med Virol ; 82(6): 1033-43, 2010 May.
Article in English | MEDLINE | ID: mdl-20419819

ABSTRACT

In 2008, measles reappeared in France in a series of outbreaks. During this period, 604 measles cases were reported to a routine surveillance system and 305 (50%) of these cases were then confirmed in the laboratory. To understand better the current epidemiological situation and the circulation of different measles strains, a phylogenetic characterization of 113 (19%) of the measles cases from these outbreaks was performed. All measles cases met the WHO clinical criteria and were confirmed either by laboratory detection of measles-specific IgM and/or by detection of the virus genome by polymerase chain reaction (PCR) and viral isolation. PCR products generated from blood, oral fluid, urine, or nasopharyngeal-swab samples were sequenced for molecular epidemiology studies. Phylogenetic analysis showed a co-circulation of genotypes D4 and D5 during the first measles outbreak in the city of Reims in early 2008. Over the course of the year, the A, B3.2, D8, and D9 genotypes also appeared. The data from this study show the simultaneous circulation of several measles genotypes in France and describe genotypes D8 and D9 for the first time in this country. The data also suggest that there are still many pockets of unvaccinated individuals helping to maintain the circulation of measles virus in the population. Phylogenetic studies allowed the corroboration of epidemiologic links and showed that nosocomial transmission can create significant risk for measles dissemination. Finally, the pattern of changes in viral genotypes during 2008 suggests a regular introduction of measles strains from abroad.


Subject(s)
Disease Outbreaks , Measles virus/classification , Measles virus/genetics , Measles/epidemiology , Measles/virology , Adolescent , Adult , Child , Child, Preschool , Cluster Analysis , Female , France/epidemiology , Genotype , Humans , Infant , Male , Measles virus/isolation & purification , Middle Aged , Molecular Epidemiology , Molecular Sequence Data , Phylogeny , RNA, Viral/genetics , Sequence Analysis, DNA , Sequence Homology , Young Adult
13.
PLoS Pathog ; 5(9): e1000587, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19806178

ABSTRACT

A number of paramyxoviruses are responsible for acute respiratory infections in children, elderly and immuno-compromised individuals, resulting in airway inflammation and exacerbation of chronic diseases like asthma. To understand the molecular pathogenesis of these infections, we searched for cellular targets of the virulence protein C of human parainfluenza virus type 3 (hPIV3-C). We found that hPIV3-C interacts directly through its C-terminal domain with STAT1 and GRB2, whereas C proteins from measles or Nipah viruses failed to do so. Binding to STAT1 explains the previously reported capacity of hPIV3-C to block type I interferon signaling, but the interaction with GRB2 was unexpected. This adaptor protein bridges Epidermal Growth Factor (EGF) receptor to MAPK/ERK pathway, a signaling cascade recently found to be involved in airway inflammatory response. We report that either hPIV3 infection or transient expression of hPIV3-C both increase cellular response to EGF, as assessed by Elk1 transactivation and phosphorylation levels of ERK1/2, 40S ribosomal subunit protein S6 and translation initiation factor 4E (eIF4E). Furthermore, inhibition of MAPK/ERK pathway with U0126 prevented viral protein expression in infected cells. Altogether, our data provide molecular basis to explain the role of hPIV3-C as a virulence factor and determinant of pathogenesis and demonstrate that Paramyxoviridae have evolved a single virulence factor to block type I interferon signaling and to boost simultaneous cellular response to growth factors.


Subject(s)
Epidermal Growth Factor/metabolism , Interferon Type I/metabolism , Parainfluenza Virus 3, Human/metabolism , Virulence Factors/metabolism , Animals , Binding Sites , Cell Count , Cell Line , Chlorocebus aethiops , Eukaryotic Initiation Factor-4E/metabolism , Flow Cytometry , GRB2 Adaptor Protein/metabolism , HeLa Cells , Humans , Immunohistochemistry , Interferon-alpha/metabolism , Interferon-beta/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Mitogen-Activated Protein Kinase Kinases/metabolism , Parainfluenza Virus 3, Human/pathogenicity , Phosphorylation , Protein Interaction Mapping , Reproducibility of Results , STAT1 Transcription Factor/metabolism , Signal Transduction , Vero Cells , Viral Proteins/metabolism
14.
BMC Infect Dis ; 9: 157, 2009 Sep 21.
Article in English | MEDLINE | ID: mdl-19772586

ABSTRACT

BACKGROUND: Serum procalcitonin (PCT) is considered useful in predicting the likeliness of developing bacterial infections in emergency setting. In this study, we describe PCT levels overtime and their relationship with bacterial infection in chronic obstructive pulmonary disease (COPD) critically ill patients with pneumonia. METHODS: We conducted a prospective cohort study in an ICU of a University Hospital. All consecutive COPD patients admitted for pneumonia between September 2005 and September 2006 were included. Respiratory samples were tested for the presence of bacteria and viruses. Procalcitonin was sequentially assessed and patients classified according to the probability of the presence of a bacterial infection. RESULTS: Thirty four patients were included. The PCT levels were assessed in 32/34 patients, median values were: 0.493 microg/L [IQR, 0.131 to 1.471] at the time of admission, 0.724 microg/L [IQR, 0.167 to 2.646] at six hours, and 0.557 microg/L [IQR, 0.123 to 3.4] at 24 hours. The highest PCT (PCTmax) levels were less than 0.1 microg/L in 3/32 (9%) patients and greater than 0.25 microg/L in 22/32 (69%) patients, suggesting low and high probability of bacterial infection, respectively. Fifteen bacteria and five viruses were detected in 15/34 (44%) patients. Bacteria were not detected in patients with PCTmax levels < 0.1 microg/L. In contrast, bacteria were detected in 4/7 (57%) patients estimated unlikely to have a bacterial infection by PCT levels (PCTmax > 0.1 and < 0.25 microg/L). CONCLUSION: Based on these results we suggest that a PCT level cut off > 0.1 microg/L may be more appropriate than 0.25 microg/L (previously proposed for non severe lower respiratory tract infection) to predict the probability of a bacterial infection in severe COPD patients with pneumonia. Further studies testing procalcitonin-based antibiotic strategies are needed in COPD patients with severe pneumonia.


Subject(s)
Calcitonin/blood , Intensive Care Units , Pneumonia, Bacterial/diagnosis , Protein Precursors/blood , Pulmonary Disease, Chronic Obstructive/complications , Aged , Aged, 80 and over , Bacteria/isolation & purification , Calcitonin Gene-Related Peptide , Female , Humans , Male , Middle Aged , Pneumonia, Bacterial/complications , Predictive Value of Tests , Prospective Studies , Pulmonary Disease, Chronic Obstructive/microbiology
15.
J Virol Methods ; 162(1-2): 119-25, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19654024

ABSTRACT

A method was developed for the detection and quantitation of HAdV (human adenovirus) and HBoV (human bocavirus) based on a duplex real-time PCR, the AB PCR, using a Smartcycler instrument. A control real-time PCR was carried out on albumin DNA to standardise the non-homogenous respiratory samples. No cross-reactivity was observed with viruses or bacteria that could be found in the respiratory tract. The diagnosis rate using the AB PCR on clinical samples was 10.7%: 3.4% for HBoV detection, 6.9% for HAdV detection and 0.3% double detection HBoV-HAdV. The clinical and epidemiological characteristics of the HAdV- and HBoV-infected patients were evaluated. In the HAdV-positive group and the HBoV-positive group the samples were classified according to the severity of the disease. The HAdV viral load did not appear to be linked to the severity of the disease. Conversely, the difference between the two HBoV groups, severe and non-severe, was significant statistically when the comparison was based on the viral load (P=0.006) or after adjustment of the viral load to the number of cells in the samples (P=0.02).


Subject(s)
Adenoviruses, Human , DNA Viruses , Human bocavirus , Polymerase Chain Reaction/methods , Respiratory Tract Infections , Adenovirus Infections, Human/diagnosis , Adenovirus Infections, Human/epidemiology , Adenovirus Infections, Human/virology , Adenoviruses, Human/classification , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , DNA Viruses/classification , DNA Viruses/genetics , DNA Viruses/isolation & purification , DNA, Viral/analysis , DNA, Viral/genetics , Female , Human bocavirus/classification , Human bocavirus/genetics , Human bocavirus/isolation & purification , Humans , Infant , Infant, Newborn , Male , Middle Aged , Parvoviridae Infections/diagnosis , Parvoviridae Infections/epidemiology , Parvoviridae Infections/virology , Reproducibility of Results , Respiratory System/virology , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/virology , Sensitivity and Specificity , Young Adult
16.
J Virol Methods ; 162(1-2): 40-5, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19638288

ABSTRACT

A 4-tube multiplex RT-PCR (mRT-PCR), which showed higher sensitivity over conventional methods, was previously developed for the diagnosis of 14 viral pathogens of the respiratory tract. Herein the mRT-PCR was compared to the commercial Luminex mPCR-microsphere flow cytometry assay (Resplex II) which allows the detection of 12 different viruses. Eleven different viruses were identified in 91 nasopharyngeal swabs of children with acute respiratory infection, influenza A (IAV) and B, respiratory syncytial virus (RSV), human rhinovirus (hRhV), human echovirus, parainfluenza viruses (PIV) 1, 2, 3 and 4, human metapneumovirus (hMPV), and human coronavirus NL63. The results of the two techniques showed 53 and 40 positive patients by the Resplex II assay and mRT-PCR, respectively, with a concordance in 35 positive and 33 negative patients (74.7%). Individual RT-PCR tests were performed to control viruses not simultaneously detected by the two multiplex assays. The major virus misdiagnosed by mRT-PCR was IAV whereas the major viruses misdiagnosed by Resplex II were PIV1, 3 and 4. The mRT-PCR remains a simple, rapid, and specific assay for the specific detection of respiratory viruses, and can be easily implemented with standards in clinical laboratories at a low cost.


Subject(s)
RNA Virus Infections , RNA Viruses/isolation & purification , Respiratory Tract Infections , Reverse Transcriptase Polymerase Chain Reaction/methods , Child , Child, Preschool , Flow Cytometry , Humans , Infant , Nasopharynx/virology , RNA Virus Infections/diagnosis , RNA Virus Infections/virology , RNA Viruses/classification , RNA Viruses/genetics , RNA, Viral/analysis , RNA, Viral/genetics , RNA, Viral/isolation & purification , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/virology , Sensitivity and Specificity , Viruses
17.
J Paediatr Child Health ; 45(3): 149-53, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19210599

ABSTRACT

AIM: The objectives of this study are to assess the frequency of human bocavirus (HBoV) infection in hospitalised children and to study the clinical symptoms associated with the detection of HBoV. METHODS: Two groups of hospitalised children were included in this study: group 1 consisted of 1946 children hospitalised from 1st September 2004 to 30th May 2005, and group 2 consisted of 448 children hospitalised from 1st November 2003 to 30th March 2004. The respiratory specimens were tested by polymerase chain reaction. RESULTS: In the first group, HBoV was detected by polymerise chain reaction in 11/828 (1.3%) of nasal specimens that tested negative for other respiratory viruses. One child tested positive for HBoV in both a nasal aspirate and stool sample. In the second group, nasal specimens were tested for all respiratory viruses, including HBoV. The presence of HBoV infection was detected in seven children (1.6%). Detection of a mixed viral population was observed in four of these children. The main symptoms in children infected with HBoV were rhinitis (50%), cough (45%), dyspnoea (28%), wheezing (28%), fever (23%) and diarrhoea (22%). The final clinical diagnoses were bronchiolitis (seven children), rhinopharyngitis (five children), the exacerbation of asthma (two children) and pneumonia (one child). Moreover, four children have associated gastroenteritis. CONCLUSION: These results contribute to the interest in the HBoV detection in children. HBoV detection in hospitalised children with or without any other respiratory virus detection was essentially associated with lower respiratory tract infection and in a lower score with upper respiratory tract infection and gastroenteritis.


Subject(s)
Bocavirus/isolation & purification , Hospitalization , Parvoviridae Infections , Base Sequence , Bocavirus/genetics , Child, Preschool , Female , Humans , Infant , Male , Polymerase Chain Reaction , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/physiopathology , Sequence Analysis
18.
PLoS One ; 4(2): e4596, 2009.
Article in English | MEDLINE | ID: mdl-19240806

ABSTRACT

BACKGROUND: RT amplification reaction has revealed that various single viruses or viral co-infections caused acute bronchiolitis in infants, and RV appeared to have a growing involvement in early respiratory diseases. Because remaining controversial, the objective was to determine prospectively the respective role of RSV, RV, hMPV and co-infections on the severity of acute bronchiolitis in very young infants. METHODS AND PRINCIPAL FINDINGS: 209 infants (median age: 2.4 months) were enrolled in a prospective study of infants <1 year old, hospitalized for a first episode of bronchiolitis during the winter epidemic season and with no high risk for severe disease. The severity was assessed by recording SaO(2)% at admission, a daily clinical score (scale 0-18), the duration of oxygen supplementation and the length of hospitalization. Viruses were identified in 94.7% by RT amplification reaction: RSV only (45.8%), RV only (7.2%), hMPV only (3.8%), dual RSV/RV (14.3%), and other virus only (2%) or coinfections (9%). RV compared respectively with RSV and dual RSV/RV infection caused a significant less severe disease with a lower clinical score (5[3.2-6] vs. 6[4-8], p = 0.01 and 5.5[5-7], p = 0.04), a shorter time in oxygen supplementation (0[0-1] days vs. 2[0-3] days, p = 0.02 and 2[0-3] days, p = 0.03) and a shorter hospital stay (3[3-4.7] days vs.6 [5-8] days, p = 0.001 and 5[4-6] days, p = 0.04). Conversely, RSV infants had also longer duration of hospitalization in comparison with RSV/RV (p = 0.01) and hMPV (p = 0.04). The multivariate analyses showed that the type of virus carried was independently associated with the duration of hospitalization. CONCLUSION: This study underlined the role of RV in early respiratory diseases, as frequently carried by young infants with a first acute bronchiolitis. RSV caused the more severe disease and conversely RV the lesser severity. No additional effect of dual RSV/RV infection was observed on the severity.


Subject(s)
Bronchiolitis/virology , Viruses/isolation & purification , Acute Disease , Bronchiolitis/pathology , Hospitalization , Humans , Infant , Infant, Newborn , Length of Stay , Metapneumovirus/isolation & purification , Prospective Studies , Respiratory Syncytial Virus, Human/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction
19.
BMC Infect Dis ; 8: 145, 2008 Oct 23.
Article in English | MEDLINE | ID: mdl-18947382

ABSTRACT

BACKGROUND: Antibiotics are recommended for severe acute exacerbation of chronic obstructive pulmonary disease (AECOPD) admitted to intensive care units (ICU). Serum procalcitonin (PCT) could be a useful tool for selecting patients with a lower probability of developing bacterial infection, but its measurement has not been investigated in this population. METHODS: We conducted a single center prospective cohort study in consecutive COPD patients admitted to the ICU for AECOPD between September 2005 and September 2006. Sputum samples or tracheal aspirates were tested for the presence of bacteria and viruses. PCT levels were measured at the time of admittance, six hours, and 24 hours using a sensitive immunoassay. RESULTS: Thirty nine AECOPD patients were included, 31 of which (79%) required a ventilator support at admission. The median [25%-75% interquartile range] PCT level, assessed in 35/39 patients, was: 0.096 microg/L [IQR, 0.065 to 0.178] at the time of admission, 0.113 microg/L [IQR, 0.074 to 0.548] at six hours, and 0.137 microg/L [IQR, 0.088 to 0.252] at 24 hours. The highest PCT (PCTmax) levels were less than 0.1 microg/L in 14/35 (40%) patients and more than 0.25 microg/L in 10/35 (29%) patients, suggesting low and high probability of bacterial infection, respectively. Five species of bacteria and nine species of viruses were detected in 12/39 (31%) patients. Among the four patients positive for Pseudomonas aeruginosa, one had a PCTmax less than 0.25 microg/L and three had a PCTmax less than 0.1 microg/L. The one patient positive for Haemophilus influenzae had a PCTmax more than 0.25 microg/L. The presence or absence of viruses did not influence PCT at time of admission (0.068 vs 0.098 microg/L respectively, P = 0.80). CONCLUSION: The likelihood of bacterial infection is low among COPD patients admitted to ICU for AECOPD (40% with PCT < 0.1 microg/L) suggesting a possible inappropriate use of antibiotics. Further studies are necessary to assess the impact of a procalcitonin-based therapeutic strategy in critically ill COPD patients.


Subject(s)
Calcitonin/blood , Intensive Care Units , Protein Precursors/blood , Pulmonary Disease, Chronic Obstructive/complications , Pulmonary Disease, Chronic Obstructive/drug therapy , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , Bacterial Infections/complications , Bacterial Infections/drug therapy , Calcitonin Gene-Related Peptide , Female , Humans , Male , Middle Aged , Predictive Value of Tests , Prospective Studies , Pulmonary Disease, Chronic Obstructive/microbiology , Sputum/microbiology , Virus Diseases/complications
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