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Biomedical subjects

Ana Avellón

Publications and source records attributed to Ana Avellón.

12 recordsLinked to original sources

Sensitivity of a modified version of the ARCHITECT Anti-HCV test in detecting samples with immunoblot-confirmed, low-level antibody to hepatitis C virus.

BACKGROUND AND OBJECTIVES: Compliance with current regulations regarding the prevention of hepatitis C virus (HCV) transmission in the blood transfusion setting requires the use of sensitive assays for HCV antibody (anti-HCV) detection, which should, ideally, identify any donor having had prior contact with the virus. Therefore, low-level anti-HCV positive blood units should be detected by the screening assays, even those reflecting a past and resolved infection. To assess the sensitivity of two versions of an automated chemiluminescent microparticle immunoassay (CMIA) for anti-HCV screening (ARCHITECT Anti-HCV), 113 single serum samples containing low levels of anti-HCV, assessed by two immunoblot tests, were selected from 3686 samples received for confirmation of HCV infection by a reference laboratory over a 2-year period. MATERIALS AND METHODS: The panel included 17 samples with HCV RNA detected by the polymerase chain reaction (PCR) and 96 PCR negative samples with either positive or indeterminate (anti-Core and anti-NS3 alone) results by immunoblot. RESULTS: All but 13 specimens (100/113, 88.5%) were detected by the current version of the ARCHITECT Anti-HCV assay and 10 additional samples (110/113, 97.3%) tested positive in a modified version of the test. CONCLUSION: The results showed that the modification introduced in the ARCHITECT Anti-HCV assay achieves a significant sensitivity improvement including samples with low-level anti-HCV which are either PCR positive or negative.

Automation↗

Frequency of hepatitis B virus 'a' determinant variants in unselected Spanish chronic carriers.

The prevalence in the population of hepatitis B virus (HBV) surface antigen (HBsAg) variants that may impair diagnosis, or allow the virus to escape vaccine-induced immunity or passive immunoglobulin therapy is unknown. A genome fragment encoding HBsAg amino acids 112-212 was amplified and sequenced from the sera of 272 unselected DNA-positive, HBV-chronic carriers from Spain. The genotype and the HBsAg subtype were predicted from the sequences. Analysis of amino-acid positions 112-157 revealed single or multiple substitutions in 39% of the carriers studied. Mutations were not detected for residues 121, 135, 137, 139, 140, 141, 142, 146, 147, 148, 149, 151, 152, 153, 155, 156, and 157. Substitutions reported previously to be in association with failures of diagnostic tests or with vaccine or immunoglobulin therapy escape were found in 12.5%, 6.6%, and 9.2% of carriers, respectively. Met133Thr (2.2%); Gln129His, Met133Ile, Phe/Tyr134Asn (1.8%); Phe/Tyr134Leu, Gly145Ala (1.5%), and Pro120Thr (1.1%) were the most frequent. Other substitutions, including Gly145Arg (0.4%), were found at a frequency of less than 1%. Samples containing HBV mutants were tested with three commercial assays for HBsAg screening. Almost all the mutants reacted to the upper cut-off values of the assays, but six samples with weak reactivity with one or more of the methods were also found. Thus, HBV mutants with a potential impact on clinical and public health issues are moderately frequent among chronic carriers from Spain, although their influence on the performance of diagnostic tests seems to be slight.

Adolescent↗

Hepatitis B virus genetic diversity.

Hepatitis B virus (HBV) is a human DNA virus, which replicates through an RNA intermediate because of the reverse-transcriptase (RT) activity of its DNA polymerase. As a result, the mutation rate for HBV is higher than the rate observed for most DNA viruses. HBVs are classified into genotypes based on genomic sequencing, and antigenic subtypes based on the antigenic properties of its major surface glycoprotein, the HBV surface antigen (HBsAg). Subgenotypes have been identified within most of the HBV genotypes. The HBV groups defined by the different genotype-HBsAg subtype associations found over the world display characteristic geographical distributions, reflecting the movements of human populations and other epidemiologically significant events. Such HBV groups constitute genetically stable viral populations sharing a common evolutionary history, but additional stable changes, originating from mutation and mutant selection, are observed within all of them. These viral sub-populations are known as the HBV variants, and some of which have medical and public health relevance. Pre-core (pre-C) defective variants have been shown to make HBV infection much less susceptible to interferon treatment, and treatment failures with other antiviral drugs have been associated with selection of resistant variants that display specific mutations in the genome region encoding the viral RT activity. Since the RT region of the genome overlaps the sequence encoding the HBsAg molecule, selection of drug resistant variants involves, in some cases, the indirect selection of HBsAg variants. Viral variants displaying changes in HBsAg seem to be very common among chronic HBV carriers; and some of these variants may emerge under the pressure of the neutralizing antibody response, leading to vaccine resistance and resistance to immunotherapy. Mutations conferring resistance to immunotherapy are noted often among liver transplant recipients and among babies born to HBV-carrier mothers. In addition, some of these HBsAg variants have been associated with lack of detection by HBsAg tests used for the diagnosis of HBV infection, for the identification of chronic carriers, for screening of blood donations for transfusion, and in the manufacture of therapeutic blood products.

Antiviral Agents↗

Acute hepatitis B virus window-period blood donations detected by individual-donation nucleic acid testing: a report on the first two cases found and interdicted in Spain.

BACKGROUND: Mathematical models predict that, in Spain, a significant number of blood units will be obtained during the window period of the hepatitis B virus (HBV) infection. Routine nucleic acid testing (NAT) on individual blood units may provide experimental data to evaluate such a theoretical risk. STUDY DESIGN AND METHODS: Between February and July 2005, a total of 34,631 individual units were screened for HBV DNA by a multiplex transcription-mediated amplification (TMA) test. Units that repeatedly reacted in the test, but did not react for HBV surface antigen (HBsAg), were submitted to additional testing by both molecular and conventional assays, and the donors were recalled for follow-up studies and the collection of clinical and epidemiologic data. RESULTS: Confirmatory testing and follow-up studies identified 2 blood units donated during the HBV infection window period (1/17,316 units studied). Sequencing of amplification products obtained by nested polymerase chain reaction (n-PCR) revealed two HBV strains from genotypes D/ayw3 and F/adw4q-, but did not identify HBsAg mutants. The HBV DNA concentration in the index donations was estimated to be below the n-PCR detection level (180 IU/mL), in both cases. One donor developed acute hepatitis 2 months after donating blood, but the other remained asymptomatic and displayed normal alanine aminotransferase levels at follow-up. CONCLUSIONS: The HBV infection window period is a real issue in the setting of Spanish blood transfusions. NAT of individual units by TMA would make a significant contribution to improving the safety of the blood supply in Spain. Additional studies involving a larger number of units and longer periods of time are required, however, to ascertain the true incidence of the problem in this country.

Acute Disease↗

Follow-up of the prevalence of hepatitis C virus genotypes in Spain during a nine-year period (1996-2004).

BACKGROUND: Recent data suggest that the prevalence of genotype 4 HCV strains among Spanish carriers is increasing. OBJECTIVE: To assess changes in the prevalence of HCV genotypes in Spain during the last nine years. METHODS: HCV RNA was amplified by the polymerase chain reaction from 3161 serum samples from unselected, anti-HCV-positive individuals, and the HCV genotype was identified by a reverse hybridisation assay (line probe assay, LiPA). Samples came from 17 different regions of Spain and were obtained between January, 1996 and December, 2004. RESULTS: The overall prevalence of HCV genotypes was: 1b, 41.3%; 1a, 24.1%; 3, 19.6%; 4, 11.6%; 2, 3.1%; and 5, 0.3%. The prevalence of genotypes 1a, 3 and 4 increased significantly among patients born after 1950, and that of genotype 1b decreased among them. These significant differences in regard to age were also observed among patients lacking notified high-risk factors. A main switch-up in prevalence of genotypes 1a and 3 was found when patients born in 1941-1950 were compared with those born in 1951-1960, but the same finding in genotype 4 was delayed to patients born in 1961-1970. CONCLUSIONS: Two separate epidemics of HCV seem to have occurred in Spain during the last 30 years. The former one involved the spread of HCV genotypes 1a and 3. The second was more recent, and involved the spread of genotype 4.

Genotype↗

[Laboratory Network within the Polio Eradication Initiative (1998-2003): six years of surveillance for acute flaccid paralysis in Spain].

INTRODUCTION: Worldwide poliomyelitis eradication was proposed in 1988 by the World Health Organization (WHO), based on reaching and maintaining high vaccination coverage and on implementing effective poliovirus infection surveillance systems. METHODS: In Spain the surveillance system focuses on active searching for acute flaccid paralysis cases through a nine-laboratory network, coordinated by the National Poliovirus Laboratory (NPL) in the National Center of Microbiology, and supported by Autonomous Community epidemiologists. Additionally, the Network sends enterovirus isolation data from other syndromes. The Laboratory Network is responsible for the primary virological study, while the NPL characterizes all polioviruses and the most epidemiologically relevant non-polio enteroviruses. RESULTS: A total of 54,533 samples were studied during the six-year period, and enteroviruses were detected in 9%. All the polioviruses isolated (n = 196), were characterized as Sabin-like (poliovirus vaccine), and among the non-polio enteroviruses, the most frequent was Echovirus 30. A total of 3% of the samples studied corresponded to the 268 acute flaccid paralysis cases or their contacts. DISCUSSION: According to the results and the WHO virological classification, Spain can be considered polio-free. However, the geographic situation of our country may facilitate the introduction of wild polioviruses that can give rise to imported poliomyelitis. Hence, the laboratory network should actively continue to participate in all the proposed WHO strategies, particularly maintenance of the poliomyelitis eradication infrastructures, and continuing monitoring and vaccination.

Humans↗

[Microbiological diagnosis of viral hepatitis].

Hepatitis of viral aetiology caused by hepatotropic virus (A, E, B, D and C) represents an important work load for the clinical virology laboratory. Most of the diagnostic is based upon detection in serum and plasma samples of different serological and virological markers, which correlates with different infection stages. In chronic infection by HBV and HCV is necessary to perform diagnostic by molecular methods as well as antigen detection in sequential samples along the course of the disease taking into account that a reliable storage must be provided for stability of structural components of the virus. Recent knowledge about mutations variants in some of the virus may alter the validity of particular markers.

Hepacivirus↗

Molecular epidemiology of hepatitis B virus in Spain: identification of viral genotypes and prediction of antigenic subtypes by limited sequencing.

The hepatitis B virus (HBV) genotypes were studied by a line probe assay (LiPA) and by direct sequencing of a 339 nucleotide fragment from the S region of the viral genome in samples from 269 carriers living in Spain, either native to Spain (231) or immigrants from Africa, Asia, and Eastern Europe (38). The sequences were also used to predict the HBV surface antigen (HBsAg) subtype on the basis of the amino acids specified at selected positions of the HBsAg molecule. Agreement between the two genotyping methods was found in most cases (98.1%) and a HBV genotype could be assigned to all samples. The viral groups D/ayw2 (30.1%), D/ayw3 (28.6%), and A/adw2 (21.2%) were prevalent, with an additional participation of the groups D/ayw4 (4.8%), F/adw4q- (1.9%), A/ayw1 (1.9%), and D/adw3 (0.7%), all of them present among the autochthonous carriers. Strains from genotypes B and C were found exclusively among Chinese immigrants. Genotype E strains were found in immigrants from Central Africa and in one patient native of Spain. Point mutations leading to amino acid changes of residues involved in the expression of the HBsAg subtype determinants were found in 12 samples (4.5%). Some mutations would predict the putative novel genotype-subtype associations A/adw4q+, A/ayr, D/ayr, and E/ayw1, while others would suggest the loss of subtype-specific determinants. The finding of HBV strains characteristic for Africa among the autochthonous carriers confirms the emergence of African HBV strains in Spain.

Adolescent↗

Quantitation of cytomegalovirus DNA in cerebrospinal fluid and serum specimens from AIDS patients using a novel highly sensitive nested competitive PCR and the cobas amplicor CMV monitor.

A novel nested quantitative-competitive polymerase chain reaction (nQC-PCR) assay was developed to quantify as few as ten copies per tube of human cytomegalovirus DNA with an overall dynamic range of 10-10(5) copies per tube. This nQC-PCR assay is based on co-amplification of a mimic DNA and it was evaluated with 26 cerebrospinal fluid (CSF) specimens and 44 serum specimens from 70 CMV-infected AIDS patients, 35 of them were diagnosed of CMV retinitis. An excellent correlation was found between nQC-PCR assay and the commercially available Cobas Amplicor CMV Monitor trade mark (CACM) assay (R = 0.9999; P < 0.001; n = 42). Moreover, 13 serum samples with CMV viral loads undetectable with the CACM were successfully quantified by nQC-PCR. CMV viral load was significantly higher in patients with CMV retinitis (P = 0.003). The nQC-PCR assay described below is a very sensitive test for accurate quantitative detection of CMV DNA in different clinical specimens that avoids the need for high-cost instrumentation.

AIDS-Related Opportunistic Infections↗

[Poliomyelitis eradication initiative. Search for and control of wild poliovirus stored at Spanish laboratories].

INTRODUCTION: An initiative to achieve global eradication of poliomyelitis was proposed in 1988. Since then, World Health Organization (WHO) efforts have been focused on reaching and maintaining high vaccination coverage and on implementing effective surveillance systems. Additionally, since 1998 the WHO has urged health authorities from countries over the world to locate and control the wild polioviruses stored in laboratories as well as potentially infectious materials that could contain these viruses or originate them under particular circumstances. METHODS: In Spain the Wild Poliovirus Containment Plan was initiated in 1999. All Spanish laboratories were inventoried and a specially designed questionnaire was distributed to them. The responses were analyzed. RESULTS: A total of 3209 laboratories were surveyed and 10 of them were found to contain infectious or potentially infectious material with wild poliovirus. The heads of these laboratories were informed about the Containment Plan and were requested to destroy this material under proper conditions. The laboratories that did not destroy the viruses adopted the BSL2/polio bio-safety measures. CONCLUSION: Despite its complexity, the search process was completed within the schedule established by the WHO, thanks to the efforts of health authorities from the Autonomous Communities. Spain reported its results to the WHO Regional Commission for the Certification of Poliomyelitis Eradication in May 2002.

Africa↗

Molecular analysis of echovirus 13 isolates and aseptic meningitis, Spain.

Echovirus 13 (EV13), considered rare, was reported worldwide in 2000, mostly related to aseptic meningitis outbreaks. In Spain, 135 EV13 isolates were identified. The genetic relationships between 64 representative strains from Spain and other reported isolates from the United States, Germany, Italy, Japan, and Sweden were described by analyzing the partial sequence of the major capsid protein (VP1) gene. The strains from Spain were clearly identified as EV13 (79.5% similarity with the EV13 reference strain) and were grouped phylogenetically into two different clusters (by origination on either the Iberian Peninsula or Canary Islands). Isolates from Germany from 2000 clustered with the Canary Islands group. The isolates from other countries obtained before 2000 were genetically distant. Changes in EV13 coding sequence involved several differences in the C-terminal extreme of the VP1 protein. Part of the neutralizing antigenic site III has been described in this genome region in poliovirus and swine vesicular disease virus.

Amino Acid Sequence↗