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1.
Transplant Proc ; 51(2): 376-379, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30879545

ABSTRACT

BACKGROUND: Hepatitis E virus (HEV) is one of the common causes of acute and chronic viral hepatitis with a global distribution. Genotypes 1 and 2 only affect humans and produce acute hepatitis epidemics in endemic regions (Asia, Africa). In nonendemic areas (America, Europe), genotypes 3 and 4 are considered a zoonosis and cause sporadic acute hepatitis. HEV has been described in solid organ transplant recipients; however, data on lung transplant patients are limited. OBJECTIVE: To present the first 3 cases of HEV infection in lung transplant recipients in our unit. CASE PRESENTATION: We report 3 cases of HEV infection in post-transplant patients presenting with symptoms and alterations in liver enzymes. All patients have no history of travel outside Spain prior to observing abnormalities in the liver function. Diagnoses were made with in-home polymerase chain reaction and enzyme-linked immunosorbent assay (IgG/IgM). The first patient was not treated and died of progressive hepatic disease, with postmortem diagnosis of HEV infection complications. The other 2 patients were treated with ribavirin after the diagnosis of HEV infection. Ribavirin was discontinued in 1 patient because of anemia necessitating red blood cell transfusions. CONCLUSIONS: HEV should be considered in the differential diagnosis of patients with abnormal liver enzymes after transplant. Early detection and treatment have implications in the prevention of liver failure and mortality. Large prospective seroprevalence studies of HEV in lung transplant patients are warranted to recognize the epidemiology of this infection in lung transplant recipients.


Subject(s)
Hepatitis E/complications , Hepatitis E/immunology , Immunocompromised Host , Lung Transplantation , Aged , Antiviral Agents/therapeutic use , Enzyme-Linked Immunosorbent Assay , Female , Hepatitis E/drug therapy , Hepatitis E virus , Humans , Male , Middle Aged , Prospective Studies , Ribavirin/therapeutic use , Transplant Recipients
2.
J Clin Virol ; 78: 20-6, 2016 May.
Article in English | MEDLINE | ID: mdl-26962723

ABSTRACT

BACKGROUND: Detection of antibodies (anti-HCV) against hepatitis C virus (HCV) is indispensable for screening and diagnosis of viral hepatitis and for the viral safety of blood, tissue or organ donations. It gains additional importance by the new HCV drugs which improve the therapeutic possibilities dramatically. OBJECTIVE: To evaluate the performance of a newly developed immune assay for anti-HCV based on the well-established VIDAS platform. STUDY DESIGN: The assay was evaluated with samples from anti-HCV negative blood donors and from patients with or without HCV markers in six centres in France, Spain and Egypt. The status of the samples was determined by using CE-marked immune assays (Architect, AxSym, Prism, Vitros), two immunoblots (RIBA, Inno-Lia) and/or HCV RNA results. RESULTS: Specificity was 99.67% in 10,320 French blood donors without anti-HCV, 99.5% in 200 anti-HCV negative hospitalized European patients and 99.0% in 198 negative patients from Egypt. Sensitivity was 99.7% in 1054 patients pretested positive by other assays; 345 patients with known genotype had genotype 1-6; 61 patients were co-infected with HIV. VIDAS was reactive in 78% of 91 patients with uncertain or very weak anti-HCV. It became on average positive at day 37 with seroconversion panels. CONCLUSIONS: This multicentric, international study with >12,000 samples show that the new VIDAS anti-HCV assay is very suitable for screening and confirmation of HCV infection. Sensitivity, specificity and recognition of seroconversion compare favorably with well-established CE-marked tests and help to clarify discrepant results obtained with other assays.


Subject(s)
Hepacivirus/immunology , Hepatitis C Antibodies/blood , Hepatitis C/diagnosis , Immunoassay/methods , Serologic Tests/methods , Animals , Egypt , France , Humans , Sensitivity and Specificity , Spain
3.
J Virol Methods ; 230: 18-23, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26784284

ABSTRACT

Hepatitis E virus (HEV) genotype 3 produces zoonotic infection associated with the consumption of infected animals. HEV infections can become chronic in immunocompromised (IC) patients. The viral genome has three well defined open reading frames (ORF1, ORF2 and ORF3) within which various domains and functions have been described. This paper (i) describes a new method of complete sequencing of the HEV coding region through overlapping PCR systems, (ii) establishes a consensus sequence and polymorphic positions (PP) for each domain, and (iii) analyzes the complete coding sequence of an IC patient. With regard to the consensus, a high percentage of PP was observed in protease (PP=19%) and the X domain (PP=22%) within ORF1, the N-terminal region of the S domain (PP=22%) in ORF2, and the P1 (PP=35%) and P2 (PP=25%) domains in ORF3. In contrast, the ORF1 Y, ORF2 S, ORF2 M and ORF3 D1 domains were conserved in the reference sequences (0.40, 1, 0.70 and 0% of PP, respectively). The sequence from the IC patient had more mutations in the RpRp (D1235G, Q1242R, S1454T, V1480I, I1502 V, K1511R, G1373 V, E1442D, V1693 M), the terminal ORF2 S- domain (F10L, S26T, G36S, S70P, A105 V, I113 V), the X domain (T938 M, T856 V, S898A) and the helicase (S1014N, S975T, Q1133 K).


Subject(s)
Genome, Viral , Genomics/methods , Hepatitis E virus/genetics , Hepatitis E/virology , Humans , Mutation , Open Reading Frames
4.
J Med Virol ; 85(11): 1975-7, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23893817

ABSTRACT

In order to investigate the etiology of viral neurological infections in Spain, a national study was performed in 2008. The results obtained have been published. Enteroviruses were the most frequent cause of the aseptic meningitis and infant febrile syndromes. The present report supplements the previous study with the genotyping of the detected enteroviruses. Typing was by amplification of partial VP1 region and sequencing in 70 (53%) of the 132 available cerebrospinal fluid samples positive for enteroviruses. Twelve different genotypes within the B species were identified. Echovirus 4 was predominant (24%), followed by echovirus 30 (19%), echovirus 9 (17%), and echovirus 6 (14%). In summary, a co-circulation of several enterovirus types associated with meningitis in children under 15 years old was observed. Although infrequently detected, echovirus 4 was the predominant genotype identified due to an aseptic meningitis outbreak which occurred in the Canary Islands in 2008.


Subject(s)
Enterovirus Infections/virology , Enterovirus/classification , Enterovirus/genetics , Meningitis, Aseptic/virology , Adolescent , Adult , Cerebrospinal Fluid/virology , Child , Child, Preschool , Enterovirus/isolation & purification , Enterovirus Infections/epidemiology , Genotype , Humans , Infant , Infant, Newborn , Male , Meningitis, Aseptic/epidemiology , Middle Aged , Prevalence , RNA, Viral/genetics , Sequence Analysis, DNA , Spain/epidemiology , Viral Structural Proteins/genetics , Young Adult
5.
J Med Virol ; 85(3): 554-62, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23239485

ABSTRACT

The aim of the study was to determine the incidence of viruses causing aseptic meningitis, meningoencephalitis, and encephalitis in Spain. This was a prospective study, in collaboration with 17 Spanish hospitals, including 581 cases (CSF from all and sera from 280): meningitis (340), meningoencephalitis (91), encephalitis (76), febrile syndrome (7), other neurological disorders (32), and 35 cases without clinical information. CSF were assayed by PCR for enterovirus (EV), herpesvirus (herpes simplex [HSV], varicella-zoster [VZV], cytomegalovirus [CMV], Epstein-Barr [EBV], and human herpes virus-6 [HHV-6]), mumps (MV), Toscana virus (TOSV), adenovirus (HAdV), lymphocytic choriomeningitis virus (LCMV), West Nile virus (WNV), and rabies. Serology was undertaken when methodology was available. Amongst meningitis cases, 57.1% were characterized; EV was the most frequent (76.8%), followed by VZV (10.3%) and HSV (3.1%; HSV-1: 1.6%; HSV-2: 1.0%, HSV non-typed: 0.5%). Cases due to CMV, EBV, HHV-6, MV, TOSV, HAdV, and LCMV were also detected. For meningoencephalitis, 40.7% of cases were diagnosed, HSV-1 (43.2%) and VZV (27.0%) being the most frequent agents, while cases associated with HSV-2, EV, CMV, MV, and LCMV were also detected. For encephalitis, 27.6% of cases were caused by HSV-1 (71.4%), VZV (19.1%), or EV (9.5%). Other positive neurological syndromes included cerebellitis (EV and HAdV), seizures (HSV), demyelinating disease (HSV-1 and HHV-6), myelopathy (VZV), and polyradiculoneuritis (HSV). No rabies or WNV cases were identified. EVs are the most frequent cause of meningitis, as is HSV for meningoencephalitis and encephalitis. A significant number of cases (42.9% meningitis, 59.3% meningoencephalitis, 72.4% encephalitis) still have no etiological diagnosis.


Subject(s)
Central Nervous System Infections/epidemiology , Central Nervous System Infections/virology , Virus Diseases/epidemiology , Virus Diseases/virology , Viruses/isolation & purification , Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Incidence , Infant , Infant, Newborn , Male , Middle Aged , Prospective Studies , Spain/epidemiology , Viruses/classification , Young Adult
6.
J Med Virol ; 84(1): 71-4, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22095537

ABSTRACT

Hepatitis E virus (HEV) is an infectious agent causing hepatitis among humans. Although hepatitis E has been reported from many European countries, its incidence in Europe is largely unknown, and the prevalence of the HEV infection is also unknown for most countries of the region. Antibody to HEV (anti-HEV) was tested on 2,305 serum samples from the general population of the Community of Madrid (Spain) collected in the year 2008 among people aged 2-60 years. Total anti-HEV was tested by enzyme-immunoassay (EIA), and reactive samples were retested separately for anti-HEV IgG and IgM by recombinant immunoblot test (RIBT). Fifty samples (2.17%) displayed reactivity for total anti-HEV after EIA testing, and anti-HEV IgG was confirmed by RIBT in 25 (1.08%). The frequency of RIBT-confirmed anti-HEV ranged from 0.97% among the youngest to 3.61% among the oldest, and displayed a statistically significant trend to increasing with age. The rate of RIBT confirmation was also significantly higher among the individuals aged above 20 years old than among those younger of 21 years. HEV infection would be less frequent in the Community of Madrid than in Catalonia or the United Kingdom, and contact with HEV would be very uncommon among children and adolescents of the region. Confirmation of EIA-reactive samples by RIBT reduced the final numbers of anti-HEV testing as much as 50%, and some findings of this study suggest that such testing protocol would reflect better the real prevalence of anti-HEV in settings of low endemicity than the single testing by EIA.


Subject(s)
Hepatitis Antibodies/blood , Hepatitis E virus/immunology , Hepatitis E/epidemiology , Adolescent , Adult , Age Factors , Child , Child, Preschool , Female , Humans , Immunoblotting , Immunoenzyme Techniques , Immunoglobulin G/blood , Immunoglobulin M/blood , Male , Middle Aged , Seroepidemiologic Studies , Spain/epidemiology , Young Adult
7.
J Clin Virol ; 50(1): 69-71, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21035388

ABSTRACT

BACKGROUND: Acute hepatitis due to hepatitis E virus (HEV) infection is both indigenous and imported to Europe. Few studies provide information about the role of HEV as an agent for acute hepatitis in Spain. OBJECTIVES: To investigate the frequency of the HEV infection among patients displaying acute hepatitis of unexplained origin in Spain, comparing the performance of two different diagnostic approaches. STUDY DESIGN: Specific IgM antibody and HEV RNA tests were used to study samples from 277 patients with acute hepatitis of unknown aetiology received during a six-year period. Samples were sent by 52 hospitals from almost all regions of Spain. RESULTS: Evidence of acute infection by HEV was obtained for 30 patients in total (10.8%), and 16 cases were unrelated to recent international travel. On samples from 158 patients tested for both anti-HEV IgM and HEV RNA at admission, the yield of IgM antibody testing (11.4%) was higher than the yield of HEV RNA testing (9.5%). CONCLUSIONS: HEV could be responsible in Spain of about 11% of cases of acute hepatitis of unknown origin overall, and of about 8% of cases unrelated to international travel or immigration. India and neighbour countries represent the highest risk for import of epidemic HEV strains into Spain. Both antibody assays and molecular tests are required to optimise the final yield of laboratory diagnosis.


Subject(s)
Antibodies, Viral , Hepatitis E virus/genetics , Hepatitis E virus/immunology , Hepatitis E/diagnosis , Molecular Diagnostic Techniques , RNA, Viral , Acute Disease , Antibodies, Viral/blood , Genotype , Humans , Molecular Diagnostic Techniques/standards , RNA, Viral/isolation & purification , Serologic Tests/standards , Spain
8.
J Clin Virol ; 47(2): 170-6, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20007023

ABSTRACT

BACKGROUND: Human enteroviruses (HEV) are the commonest cause of viral meningitis as well as other pathologies, therefore HEV characterization is important both in patient management and epidemiological investigation. OBJECTIVES: A 10-year study of patients with enteroviral infection was carried out in Spain to determine the underlying etiology. STUDY DESIGN: HEV were fully typed by microneutralisation tests and/or molecular methods. RESULTS: A collection of 86404 clinical samples were studied in several Spanish laboratories. These were collected from patients with different syndromes, mainly aseptic meningitis (AM), fever, respiratory diseases and acute flaccid paralysis. Of these, 6867 HEV were obtained. At the National Poliovirus Laboratory 2814 were serotypically characterised. Among non-polio enteroviruses, the eight main serotypes were Echovirus 30 (25%), Echovirus 6 (12.4%), Echovirus 13 (8.3%), Echovirus 11 (7.4%) and Echovirus 9 (4.7%), followed by Coxsackievirus B5 (4.2%) and Echovirus 7 and Coxsackievirus A9 (3.7%) each. In AM cases, Echovirus 30 was identified in 39% of them, followed by Echovirus 6 (14%). However, Echovirus 6 was mainly associated with respiratory disease (17%), followed by Echovirus 11 (10%). On the other hand, Echovirus 30, Echovirus 11 and Echovirus 6 contributed equally with 12% of each serotype in the cases of fever. CONCLUSIONS: The present report complements previous data (Trallero et al.(13)), with the results of HEV incidence in Spain from 1998 to 2007. The surveillance described in this study provided valuable information as to which serotypes are in circulation, the emergence of new HEV and association with clinical manifestations.


Subject(s)
Enterovirus Infections/epidemiology , Enterovirus Infections/virology , Enterovirus/classification , Enterovirus/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Genotype , Humans , Incidence , Infant , Infant, Newborn , Male , Middle Aged , Neutralization Tests , Sequence Analysis, DNA , Serotyping , Spain/epidemiology , Young Adult
9.
J Med Virol ; 81(10): 1743-9, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19697406

ABSTRACT

Hepatitis E virus (HEV) causes hepatitis E, an acute liver disease displaying diverse epidemiological patterns that correlate with the genetic diversity of the virus. Only a few strains have been characterized to date from cases of hepatitis E in Spain. Using three sets of new, HEV-specific primers, viral genome fragments were amplified from serum samples from 13 patients with acute hepatitis in different regions of Spain. Direct sequencing of these fragments and analysis of sequences lead to identify six genotype 1, six genotype 3, and one genotype 4 viral strains. Genotype 1 sequences were found in the clade with subtype 1a strains, and were amplified from travelers from India and Bangladesh, and from an African immigrant. Genotype 3 sequences were found in the clade with subtype 3f strains, were always amplified from patients who did not travel abroad recently, and were closely related to sequences from swine strains isolated in Spain. Patients infected by these strains lived in different regions and were mainly men aged above 50 years. The single genotype 4 sequence detected was amplified from a traveler returning from Vietnam. Hepatitis E is both an imported and an autochthonous disease in Spain, and closely related HEV genotype 3f strains are responsible for infections acquired locally in different regions of the country within a given time. Studies involving a significant number of human, swine, and environmental viral strains collected prospectively are, however, required in order to confirm a swine origin for autochthonous HEV genotype 3 human infections.


Subject(s)
Hepatitis E virus/isolation & purification , Hepatitis E/epidemiology , Hepatitis E/virology , Adolescent , Adult , Aged , Animals , Cluster Analysis , DNA Primers/genetics , Emigrants and Immigrants , Female , Genotype , Humans , Male , Middle Aged , Molecular Epidemiology , Molecular Sequence Data , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Sequence Analysis, DNA , Sequence Homology , Serum/virology , Spain/epidemiology , Travel , Zoonoses/virology
10.
J Clin Virol ; 45(3): 259-61, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19505848

ABSTRACT

BACKGROUND: The accuracy of the diagnosis of hepatitis E in the clinical setting relies mainly on the performance of assays for hepatitis E virus (HEV)-specific IgM (anti-HEV IgM) testing in serum. OBJECTIVES: Identification of factors influencing the specificity of the results obtained with these assays is an important issue in regard to the accuracy of the diagnosis. STUDY DESIGN: Anti-HEV IgM and HEV RNA were studied in samples from 153 patients with acute hepatitis of unknown aetiology received during a two-year period. Fifteen patients were positive for anti-HEV IgM, and eight of them were also positive for HEV RNA. Investigation of CMV and Epstein-Barr virus (EBV) infection markers among the remaining seven patients, and of HEV infection markers among 18 patients with infectious mononucleosis, was performed. RESULTS: The results obtained showed that acute infection by CMV or EBV may cause false reactivity for anti-HEV IgM, likely because of polyclonal B-cell stimulation. CONCLUSIONS: Since infection by these herpesviruses may produce acute hepatitis, such event can cause diagnostic mistakes and should be investigated in patients positive for anti-HEV IgM and negative for HEV RNA.


Subject(s)
Cytomegalovirus Infections/diagnosis , Epstein-Barr Virus Infections/diagnosis , Hepatitis E/diagnosis , Adolescent , Adult , Child , Child, Preschool , Diagnosis, Differential , False Positive Reactions , Female , Hepatitis Antibodies/blood , Hepatitis E virus/genetics , Hepatitis E virus/immunology , Hepatitis E virus/isolation & purification , Humans , Immunoglobulin M/blood , Infant , Infant, Newborn , Middle Aged , Pregnancy , RNA, Viral/blood , Young Adult
11.
J Appl Microbiol ; 105(6): 1945-55, 2008 Dec.
Article in English | MEDLINE | ID: mdl-19120641

ABSTRACT

AIMS: Update information regarding occurrence and levels of culturable enteroviruses in several types of surface polluted waters in north-eastern Spain and determine the proportion of the different species and serotypes. METHODS AND RESULTS: The best procedures on hand in our laboratory for concentrating and quantifying culturable enteroviruses from different water sample types were used. Sequencing was used for typing the virus isolates. Geometric means of enteroviruses densities expressed in plaque forming units per litre were 968 in raw sewage, 12.51 in secondary effluents, 0.017 in tertiary effluents, 0.4 in river water and 0.36 in seawater. Enterovirus densities in wastewater revealed certain seasonality with a maximum at the end of spring - beginning of the summer. Coxsackievirus B, and amid them serotype CB4, were the most abundant species and serotypes detected. CONCLUSIONS: Densities of enteroviruses in different north-eastern Spain surface waters are similar to those present in industrialized countries with temperate climate. No wild polioviruses were detected. Distribution of species showed a clear prevalence of coxsackieviruses. SIGNIFICANCE AND IMPACT OF THE STUDY: Information regarding enteroviruses in this geographical area provides valuable information to estimate the risk of enteroviruses transmission through water and for complementing clinical epidemiological data.


Subject(s)
Enterovirus/isolation & purification , Water Microbiology , Water Pollution , Animals , Colony Count, Microbial , Enterovirus/classification , Environmental Monitoring/methods , Fresh Water , Seasons , Seawater , Serotyping , Sewage , Spain , Waste Disposal, Fluid
12.
Clin Microbiol Infect ; 13(4): 369-76, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17359320

ABSTRACT

Pre- vs. post-vaccination changes in correlations between IgG concentrations (ELISA titres) and opsonophagocytic activity (OPA) against Streptococcus pneumoniae serotypes 6B, 14 and 23F induced by the 23-valent polysaccharide vaccine were studied in paired serum samples received from elderly individuals, haemodialysed patients and kidney transplant recipients by the Spanish Pneumococcal Reference Laboratory. The pre- and post-vaccination parameters considered were: ELISA and OPA titres and the percentage of subjects with post-vaccination OPA values above the cut-off levels; the correlations between OPA and ELISA (Spearman correlation coefficient, r); and the amount of IgG needed to obtain OPA (beta coefficient). Non-significant pre-vaccination correlations between OPA and ELISA were found. Vaccination increased the correlation coefficient between OPA and ELISA to a statistically significant level for serotypes 6B, 14 and 23F in samples from haemodialysed patients, for serotypes 14 and 23F in samples from elderly individuals, and for none of the serotypes in samples from transplant recipients. In all cases, except for serotype 23 in transplant recipients, vaccination increased the beta coefficient, indicating that lower amounts of IgG were needed to obtain high OPA titres. A globally lower response was obtained for serotype 23 and/or transplant recipients.


Subject(s)
Antibodies, Bacterial/blood , Phagocytosis , Pneumococcal Vaccines/immunology , Streptococcus pneumoniae/immunology , Vaccination , Adult , Aged , Enzyme-Linked Immunosorbent Assay , Humans , Immunoglobulin G/blood , Middle Aged , Morbidity , Serotyping , Streptococcus pneumoniae/classification
13.
J Virol Methods ; 135(2): 281-7, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16713633

ABSTRACT

The Lyssavirus genus includes seven species or genotypes named 1-7. Rabies genotypes correlate with geographical distribution and specific hosts. Co-circulation of different lyssaviruses, imported cases, and the presence of unknown viruses, such as Aravan, Khujand, Irkut and West Caucasian Bat Virus, make it necessary to use generic methods able to detect all lyssaviruses. Primer sequences were chosen from conserved regions in all genotypes in order to optimise a generic RT-PCR. Serial dilutions of 12 RNA extracts from all seven Lyssavirus genotypes were examined to compare the sensitivity of the RT-PCR standardised in this study with a published RT-PCR optimised for EBLV1 detection and capable of amplifying RNA from all seven lyssaviruses. All seven genotypes were detected by both RT-PCRs, however, the sensitivity was higher with the new version of the test. Twenty samples submitted for rabies diagnosis were tested by the new RT-PCR. Eight out of 20 samples from six dogs, one horse and one bat were found positive, in agreement with immunofluorescence results. Seven samples from terrestrial mammals were genotype 1 and one from a bat was genotype 5. In conclusion, this method can be used to complement immunofluorescence for the diagnosis of rabies, enabling the detection of unexpected lyssaviruses during rabies surveillance.


Subject(s)
Dogs/virology , Lyssavirus/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Animals , Chiroptera/virology , Fluorescent Antibody Technique , Genotype , Horses/virology , Lyssavirus/classification , Lyssavirus/isolation & purification , RNA, Viral/analysis , Rabies/diagnosis
14.
J Clin Microbiol ; 43(12): 6176-82, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16333124

ABSTRACT

Here we present a system for adenovirus detection and genotyping based on PCR amplification and phylogenetic analysis of a conserved hexon gene fragment. The system was validated using 157 sequences (86 previously typed and 71 clinical samples) and correctly identified species and serotype in 100% and 84% of sequences, respectively. Known associations between specific serotypes and clinical syndromes are verified. Possible new associations are described to allow further independent testing.


Subject(s)
Adenovirus Infections, Human/virology , Adenoviruses, Human/classification , Capsid Proteins/genetics , Polymerase Chain Reaction/methods , Sequence Analysis, DNA , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Adolescent , Adult , Child , Child, Preschool , DNA, Viral/analysis , Genome, Viral , Humans , Infant , Infant, Newborn , Middle Aged , Molecular Sequence Data , Phylogeny , Serotyping
15.
Epidemiol Infect ; 130(2): 251-6, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12729193

ABSTRACT

Echoviruses are the commonest cause of aseptic meningitis (AM). Echovirus type 13 (EV-13) was the second enterovirus serotype associated with different local outbreaks of AM in Spain between February and October 2000. It was the first time that an epidemic AM caused by this virus was recognized in Spain. The index case appeared in the Canary Islands (Canarias). The EV-13 virus was isolated from 135 patients, predominantly from cerebrospinal fluid (CSF). All isolates were from children under 13 years. The age specific peak incidence was in infants under 1 year. Most patients had fever, headache and other meningeal signs. This enterovirus serotype, not previously detected in Spain, caused severe illness with a high attack rate.


Subject(s)
Disease Outbreaks , Echovirus Infections/epidemiology , Meningitis, Aseptic/epidemiology , Adolescent , Age Distribution , Child , Child, Preschool , Echovirus Infections/cerebrospinal fluid , Female , Humans , Infant , Infant, Newborn , Male , Meningitis, Aseptic/cerebrospinal fluid , Spain/epidemiology
16.
J Clin Microbiol ; 39(10): 3678-83, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11574590

ABSTRACT

Brain analysis cannot be used for the investigation of active lyssavirus infection in healthy bats because most bat species are protected by conservation directives. Consequently, serology remains the only tool for performing virological studies on natural bat populations; however, the presence of antibodies merely reflects past exposure to the virus and is not a valid marker of active infection. This work describes a new nested reverse transcription (RT)-PCR technique specifically designed for the detection of the European bat virus 1 on oropharyngeal swabs obtained from bats but also able to amplify RNA from the remaining rabies-related lyssaviruses in brain samples. The technique was successfully used for surveillance of a serotine bat (Eptesicus serotinus) colony involved in a case of human exposure, in which 15 out of 71 oropharyngeal swabs were positive. Lyssavirus infection was detected on 13 oropharyngeal swabs but in only 5 brains out of the 34 animals from which simultaneous brain and oropharyngeal samples had been taken. The lyssavirus involved could be rapidly identified by automatic sequencing of the RT-PCR products obtained from 14 brains and three bat oropharyngeal swabs. In conclusion, RT-PCR using oropharyngeal swabs will permit screening of wild bat populations for active lyssavirus infection, for research or epidemiological purposes, in line not only with conservation policies but also in a more efficient manner than classical detection techniques used on the brain.


Subject(s)
Chiroptera/virology , Lyssavirus/isolation & purification , Oropharynx/virology , RNA, Viral/analysis , Rhabdoviridae Infections/veterinary , Animals , Animals, Wild , Brain/virology , Humans , Lyssavirus/classification , Lyssavirus/genetics , Reverse Transcriptase Polymerase Chain Reaction , Rhabdoviridae Infections/diagnosis , Rhabdoviridae Infections/virology
17.
J Virol Methods ; 92(2): 113-20, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11226558

ABSTRACT

A new adenovirus specific nested polymerase chain reaction (PCR) method is described. It was designed inside the hexon protein gene of the adenovirus genome, and was able to detect DNA of all 47 human adenovirus types in a wide range of clinical samples. A sensitive internal control system able to assure proper analytical conditions for the amplification of as few as 100 molecules of a heterologous DNA was included to avoid false negative results. Sensitivity was estimated at about 10 molecules per tube of a plasmid containing an insert of the first amplification product. The method was able to detect adenovirus infection in 31/43 conjunctival scrapings from patients with acute kerato conjunctivitis 10/40 nasopharyngeal aspirates from patients admitted to hospital with acute respiratory disease and 2/26 urine samples from patients with haemorrhagic cystitis with better sensitivity than cell culture or rapid diagnosis by antigen detection by immunofluorescence (IF) in the case of respiratory specimens. Only two of 17 stools positive for a group F adenovirus specific latex immunoassay were PCR negative. The internal control system avoided a false negative result on another two stool samples. In conclusion, the method described below was shown to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection by IF.


Subject(s)
Adenovirus Infections, Human/diagnosis , Adenoviruses, Human/isolation & purification , DNA, Viral/analysis , Polymerase Chain Reaction/methods , Adenovirus Infections, Human/virology , Adenoviruses, Human/classification , Adenoviruses, Human/genetics , Adult , Antigens, Viral/analysis , Cell Line , Child , Child, Preschool , Humans , Reference Standards , Sensitivity and Specificity , Virus Cultivation
18.
J Virol Methods ; 88(1): 51-61, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10921842

ABSTRACT

A new method to quantitate small amounts of DNA in clinical specimens is described. The method, a nested competitive polymerase chain reaction (ncPCR), is able to quantitate between 10 and 10(6) copies per tube of polyomavirus DNA and shows good reproducibility when clinical samples are analysed. Throughout the whole procedure, an internal standard (IS) competes for the primers with the target DNA. The internal standard, a heterologous sequence containing the four primer recognition sites, was constructed using a modification of the 'MIMIC' approach that is useful for obtaining competitor sequences for any viral, bacterial or eukaryotic target. The ncPCR method for polyomavirus was applied to cerebrospinal fluids (CSF) from AIDS patients with progressive multifocal leukoencephalopathy (PML) and urine specimens from bone marrow transplant patients affected by haemorrhagic cystitis. The results obtained suggest that the ncPCR method is a sensitive and useful method for quantitating genomic load in clinical samples.


Subject(s)
Acquired Immunodeficiency Syndrome/complications , Cystitis/virology , DNA, Viral/analysis , JC Virus/isolation & purification , Leukoencephalopathy, Progressive Multifocal/virology , Polymerase Chain Reaction/methods , Bone Marrow Transplantation/adverse effects , Cerebrospinal Fluid/virology , Hemorrhage , Humans , JC Virus/genetics , Sensitivity and Specificity , Urine/virology , Viral Load
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