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1.
Cerebrovasc Dis ; 50(6): 707-714, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34175851

RESUMEN

OBJECTIVE: To describe the impact of COVID-19 on acute cerebrovascular disease care across 9 comprehensive stroke centers throughout Los Angeles County (LAC). METHODS: Volume of emergency stroke code activations, patient characteristics, stroke severity, reperfusion rates, treatment times, and outcomes from February 1 to April 30, 2020, were compared against the same time period in 2019. Demographic data were provided by each participating institution. RESULTS: There was a 17.3% decrease in stroke code activations across LAC in 2020 compared to 2019 (1,786 vs. 2,159, respectively, χ2 goodness of fit test p < 0.0001) across 9 participating comprehensive stroke centers. Patients who did not receive any reperfusion therapy decreased by 16.6% in 2020 (1,527) compared to 2019 (1,832). Patients who received only intravenous thrombolytic (IVT) therapy decreased by 31.8% (107 vs. 157). Patients who received only mechanical thrombectomy (MT) increased by 3% (102 vs. 99). Patients who received both IVT and MT decreased by 31.8% (45 vs. 66). Recanalization treatment times in 2020 were comparable to 2019. CSCs serving a higher proportion of Latinx populations in the eastern parts of LAC experienced a higher incidence of MT in 2020 compared to 2019. Mild increase in stroke severity was seen in 2020 compared to 2019 (8.95 vs. 8.23, p = 0.046). A higher percentage of patients were discharged home in 2020 compared to 2019 (59.5 vs. 56.1%, p = 0.034), a lower percentage of patients were discharged to skilled nursing facility (16.1 vs. 20.7%, p = 0.0004), and a higher percentage of patients expired (8.6 vs. 6.3%, p = 0.008). CONCLUSION: LAC saw a decrease in overall stroke code activations in 2020 compared to 2019. Reperfusion treatment times remained comparable to prepandemic metrics. There has been an increase in severe stroke incidence and higher volume of thrombectomy treatments in Latinx communities within LAC during the pandemic of 2020. More patients were discharged home, less patients discharged to skilled nursing facilities, and more patients expired in 2020, compared to the same time frame in 2019.


Asunto(s)
Isquemia Encefálica/epidemiología , COVID-19 , Fibrinolíticos/efectos adversos , Accidente Cerebrovascular Isquémico , Accidente Cerebrovascular/terapia , Terapia Trombolítica , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/terapia , Humanos , Los Angeles/epidemiología , Estudios Retrospectivos , SARS-CoV-2 , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/epidemiología , Trombectomía , Tiempo de Tratamiento , Resultado del Tratamiento
2.
J Neurosci Nurs ; 50(1): 22-23, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29266083

RESUMEN

A collaborative approach was used to ascertain an appropriate stimulus for the patients to remember their stroke-specific education. The stroke education had to stand out amidst the myriad of papers and folders patients are bombarded with in the hospital. The team came up with the simple idea of using a bright red folder. When the patients were called, the call center would prompt the patient by saying, "The stroke education was given to you in a bright red folder." Before the implementation of the red folders, only 81.5% of the patients remembered receiving stroke education. After the implementation of the red folders, 96.8% remembered receiving stroke education. The principle of Occam's razor proved to be correct in our study. A very simple idea such as changing the color of the folders to bright red proved to have very meaningful results.


Asunto(s)
Educación del Paciente como Asunto , Accidente Cerebrovascular , Conductas Relacionadas con la Salud , Humanos
3.
Hepatol Res ; 38(7): 704-16, 2008 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-18328069

RESUMEN

AIM: To study hepatitis C virus (HCV) selection and hypervariable region-1 (HVR1) evolution in a chimpanzee chronically infected with HCV-1 over 12 years after inoculation with a human factor VIII concentrate contaminated with HCV. METHODS: From the inoculum, the earliest chimpanzee plasma and 12 annual plasma samples, HCV fragments including HVR1 were amplified followed by cloning and sequencing. RESULTS: Five HCV subtypes - 1a, 1b, 2a, 2b, 3a - and multiple 1a strains were identified in the inoculum. Two 1a strains were found in the earliest chimpanzee sample, while a single HCV-1 strain was detected in the 12 annual samples. None of the chimpanzee sequences were identical to those found in the inoculum. Over 12 years, HVR1 patterns changed irregularly, but a few patterns showed identical nucleotide or amino acid sequences. In the last three years, the variety of HVR1 patterns decreased, while the proportion of major patterns increased. These corresponded to a higher virus load and a lower number of amino acid substitutions. Simultaneously, the HVR1 sequences became more similar to the consensus sequence of the 1a subtype. CONCLUSION: HCV selection was observed from the inoculum to the inoculated chimpanzee and from the early acute hepatitis to the persistent chronic infection. The selection occurred at three levels: among subtypes after transmission, among isolates during acute hepatitis and among quasispecies in chronic infection.

4.
J Med Virol ; 79(11): 1696-702, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17854031

RESUMEN

Large outbreaks and sporadic cases of hepatitis E have been reported in Central Asia. We assessed the genetic relatedness of hepatitis E virus (HEV) strains from outbreak and sporadic cases in Turkmenistan. Specimens from outbreak and sporadic cases of acute hepatitis non-A, non-B were tested by reverse transcription (RT)-polymerase chain reaction (PCR) to identify the presence of HEV RNA; nucleotide sequences were analyzed. HEV RNA was detected from 23/156 (15%) outbreak cases and 2/23 (9%) sporadic cases. The HEV outbreak isolates represented 14 unique sequences with genetic distances varying between 0.3% and 8.6%, 12 of which were closely related, with distances between 0.3% and 5.6%. Two unique sequences from outbreak cases 32 and 42 were closely related (99.7%) and shared 91.8-93.4% of sequence with the other 12 strains. The two strains were closely related to the previously published isolates from Burma (99.7-100%) and India-Madras (95.7-96.1%). The two 1994 sporadic HEV strains were 97.4% distinct, wile revealing 91.4-94.1% homology to 1985 strains, and 94.4-94.7% to HEV from the neighboring China and Pakistan. Genetic diversity of HEV that caused the hepatitis E outbreak in Turkmenistan in 1985 suggests heterogeneity of viral sources. Sporadic hepatitis E that occurred in 1994 was caused by viral strains genetically distinct from those causing the outbreak in 1985, yet closely related to HEV from neighboring countries. The study suggests that circulation of a broad variety of strains of HEV may occur in Central Asia, regardless of international borders, presenting a significant public health threat to the population of the region.


Asunto(s)
Brotes de Enfermedades , Virus de la Hepatitis E/clasificación , Virus de la Hepatitis E/genética , Hepatitis E/epidemiología , Hepatitis E/virología , Enfermedad Aguda , Adulto , Femenino , Virus de la Hepatitis E/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Sistemas de Lectura Abierta/genética , Embarazo , Complicaciones Infecciosas del Embarazo/epidemiología , Complicaciones Infecciosas del Embarazo/virología , ARN Viral/sangre , ARN Viral/aislamiento & purificación , Análisis de Secuencia de ADN , Turkmenistán/epidemiología
5.
J Gen Virol ; 87(Pt 3): 629-634, 2006 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-16476984

RESUMEN

Here, the complete genome sequences for three hepatitis C virus (HCV) variants identified from China and belonging to genotype 6 are reported: km41, km42 and gz52557. Their entire genome lengths were 9430, 9441 and 9448 nt, respectively; the 5' untranslated regions (UTRs) contained 341, 342 and 339 nt, followed by single open reading frames of 9045, 9045 and 9057 nt, respectively; the 3' UTRs, up to the poly(U) tracts, were 41, 51 and 52 nt, respectively. Phylogenetic analyses showed that km41 is classified into subtype 6k and km42 into subtype 6n. Although gz52557 clustered distantly with subtype 6g, it appeared to belong to a distinct subtype. Analysis with 53 and 105 partial core and NS5B region sequences, respectively, representing 17 subtypes from 6a to 6q and three unassigned isolates of genotype 6 in co-analyses demonstrated that gz52557 was equidistant from all of these isolates, indicating that it belongs to a novel subtype. However, based on a recent consensus that three or more examples are required for a new HCV subtype designation, it is suggested that gz52557 remains unassigned to any subtype.


Asunto(s)
Hepacivirus/clasificación , China , Genoma Viral , Hepacivirus/genética , Hepatitis C/virología , Humanos , Datos de Secuencia Molecular , Especificidad de la Especie
6.
J Virol Methods ; 131(1): 65-71, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16125257

RESUMEN

Hepatitis E virus (HEV) is transmitted by the fecal-oral route and causes sporadic and epidemic forms of acute hepatitis. Large waterborne HEV epidemics have been documented exclusively in developing countries. At least four major genotypes of HEV have been reported worldwide: genotype 1 (found primarily in Asian countries), genotype 2 (isolated from a single outbreak in Mexico), genotype 3 (identified in swine and humans in the United States and many other countries), and genotype 4 (identified in humans, swine and other animals in Asia). To better detect and quantitate different HEV strains that may be present in clinical and environmental samples, we developed a rapid and sensitive real-time RT-PCR assay for the detection of HEV RNA. Primers and probes for the real-time RT-PCR were selected based on the multiple sequence alignments of 27 sequences of the ORF3 region. Thirteen HEV isolates representing genotypes 1-4 were used to standardize the real-time RT-PCR assay. The TaqMan assay detected as few as four genome equivalent (GE) copies of HEV plasmid DNA and detected as low as 0.12 50% pig infectious dose (PID50) of swine HEV. Different concentrations of swine HEV (120-1.2PID50) spiked into a surface water concentrate were detected in the real-time RT-PCR assay. This is the first reporting of a broadly reactive TaqMan RT-PCR assay for the detection of HEV in clinical and environmental samples.


Asunto(s)
Virus de la Hepatitis E/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Animales , Cartilla de ADN , Virus de la Hepatitis E/genética , ARN Viral/genética , Sensibilidad y Especificidad , Porcinos/virología , Proteínas Virales , Microbiología del Agua
7.
J Gen Virol ; 87(Pt 1): 73-82, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16361419

RESUMEN

Subtype 1b is the most common strain of Hepatitis C virus (HCV) in China. Here, the molecular epidemiology and epidemic history of this strain were investigated by conducting phylogenetic and population genetic analyses of E1 and NS5B gene sequences sampled from nine Chinese cities. The phylogenetic analysis indicated the presence of two clusters of Chinese strains that did not include reference strains from other countries, suggesting that these clusters represent two independent chains of HCV transmission within China. The remaining Chinese isolates were more closely related to reference strains from other countries. The date of origin and past population dynamics of the two groups were investigated using a new population genetic method, the Bayesian skyline plot. The estimated dates of origin of both groups coincide with the period of the Chinese 'Cultural Revolution' during the years 1966-1976. Both groups grew at a rapid exponential rate between approximately 1970 and approximately 1990, after which transmission slowed considerably. Possible explanations for the groups' fast spread and subsequent slowdown are discussed, including parenteral transmission by unsafe injection, iatrogenic transmission by infected blood or blood products and improvements in blood safety since 1990. These results shed light on HCV transmission in China and may help to predict the future burden of HCV-related disease in the country.


Asunto(s)
Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C/epidemiología , Epidemiología Molecular , China/epidemiología , ADN Viral/análisis , Hepatitis C/transmisión , Hepatitis C/virología , Hepatitis Viral Humana/epidemiología , Hepatitis Viral Humana/virología , Humanos , Filogenia
8.
J Gen Virol ; 87(Pt 1): 83-91, 2006 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16361420

RESUMEN

This study determined whether selective transmission of hepatitis C virus (HCV) species occurred among human and chimpanzee recipients of contaminated blood products or plasma containing multiple genotypes, subgenotypes and quasispecies. Commercially prepared factor VIII concentrate (lot DO56), produced prior to HCV testing and inactivation, was subsequently found by direct cloning to contain the following subgenotypes: 1a and 1b (73 % of clones), 2a (13 % of clones), 2b (11 % of clones) and 3a (4 % of clones). A patient transfused with factor VIII concentrate DO56 was diagnosed with clinical non-A, non-B hepatitis and subsequently found to be infected with HCV subgenotype 1b. Among five chimpanzees inoculated experimentally with the same factor VIII concentrate, two were infected only with HCV subgenotype 1a and three were infected with approximately equivalent clonal proportions of subgenotypes 1a and 1b. HCV hypervariable region 1 (HVR1) quasispecies analysis of the DO56 factor VIII concentrate and a serum specimen from the single chimpanzee that developed a chronic HCV infection following inoculation with DO56 showed 0-56 % nucleotide variation. However, specimens from chimpanzees infected in the second to fourth passages of the DO56 inoculum had 0-8 % HVR1 quasispecies nucleotide variation. The high HVR1 quasispecies variation in the factor VIII concentrate and its first passage in chimpanzees indicates the presence of multiple HCV isolates, whereas the low variation in the second to fourth chimpanzee passages suggests transmission of a single HCV isolate. These findings strongly suggest selective transmission of HCV isolates during experimental chimpanzee infection and among humans exposed to multiple HCV species.


Asunto(s)
Factor VIII/administración & dosificación , Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C/transmisión , Reacción a la Transfusión , Proteínas del Envoltorio Viral/genética , Animales , Modelos Animales de Enfermedad , Genotipo , Hepacivirus/aislamiento & purificación , Hepatitis C/virología , Humanos , Datos de Secuencia Molecular , Pan troglodytes , Proteínas del Envoltorio Viral/sangre , Proteínas del Envoltorio Viral/metabolismo
9.
J Virol Methods ; 126(1-2): 139-48, 2005 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-15847930

RESUMEN

The goal of this study was to adapt a long RT-PCR technique to amplify large PCR fragments from the genome of hepatitis C virus (HCV) isolates using clinical samples. This was done by using a reverse transcriptase devoid of RNase H activity and a mixture of two antibody-bound thermostable polymerases to combine the high processivity of Taq and the high fidelity of Pwo with its 3'-->5' exonuclease activity. Other modifications included gentle handling during RNA extraction, the absence of tRNA and random primers, a two-step reverse transcription procedure to optimize cDNA synthesis, and increasing the annealing temperature for primers. With this approach, the HCV-1 genome (nucleotides 35-9282) was amplified consistently as two overlapping fragments of 5344 and 4675 bp from a pooled chimpanzee plasma sample containing approximately 10(6) genome copies of HCV RNA/ml. Using the conditions that we identified, 96% of the complete genomic sequence of a distinct HCV genotype 6 variant (km45) was determined from less than 300 microl of serum. This method should prove useful for molecular, epidemiological and clinical studies of hepatitis C where samples are limited but complete virus sequence is required, for example, identifying mutational hot spots of HCV under specific clinical conditions.


Asunto(s)
Genoma Viral , Hepacivirus/genética , Hepatitis C/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Animales , Secuencia de Bases , ADN Polimerasa Dirigida por ADN/metabolismo , Hepacivirus/clasificación , Datos de Secuencia Molecular , Pan troglodytes , Filogenia , Plasma/virología , Homología de Secuencia de Ácido Nucleico , Polimerasa Taq/metabolismo
10.
Annu Rev Public Health ; 26: 281-302, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-15760290

RESUMEN

The realm of diagnostic assays for detection of acute infections is rapidly changing from antibody detection to pathogen detection, from clinical laboratory based to point-of-care based, from single analyte detection to multiple analyte detection, and is more focused on detection using less invasive approaches for collecting biological samples. New assays are typically more sensitive than are conventional assays and have the capability of providing more information that characterizes the pathogen or the host response to the pathogen. From a public health perspective, the advent of molecular epidemiology, which allows tracking of pathogens based on unique genetic sequences or antigenic properties, has revolutionized how epidemiologists investigate and evaluate epidemics and assess endemic diseases. In addition, the use of point-of-care (POC) devices can impact the detection and surveillance of infections and will enhance our ability to accurately identify the causes of illnesses.


Asunto(s)
Enfermedades Transmisibles/diagnóstico , Enfermedades Transmisibles/microbiología , Técnicas Microbiológicas/métodos , Salud Pública/métodos , Causalidad , Control de Enfermedades Transmisibles/métodos , Enfermedades Transmisibles/epidemiología , Citometría de Flujo/métodos , Predicción , Humanos , Técnicas para Inmunoenzimas/métodos , Análisis por Micromatrices/métodos , Técnicas Microbiológicas/normas , Técnicas Microbiológicas/tendencias , Técnicas de Diagnóstico Molecular/métodos , Epidemiología Molecular/métodos , Sistemas de Atención de Punto , Reacción en Cadena de la Polimerasa/métodos , Vigilancia de la Población/métodos , Salud Pública/normas , Salud Pública/tendencias , Sensibilidad y Especificidad , Manejo de Especímenes/métodos , Evaluación de la Tecnología Biomédica , Estados Unidos
11.
J Med Virol ; 75(4): 538-49, 2005 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-15714489

RESUMEN

To determine hepatitis C virus (HCV) genotype distribution in China, a total of 148 HCV RNA positive serum samples were collected from nine geographic areas and subjected to RT-PCR followed by direct DNA sequencing and phylogenetic analysis of the core, E1, and NS5B regions. HCV was genotyped in 139 (93.9%) samples. Among them subtype 1b was the most predominant [66% (92/139)] followed by 2a [14% (19/139)]. Of 92 subtype 1b isolates, 35 (38%) and 30 (33%) formed two clusters, designated groups A and B. Group A was prevalent throughout China, while group B was predominant in the central and southern regions. In three cities in the Pearl River Delta, subtype 6a replaced 2a as the second most predominant subtype, and in Kunming (southwest) multiple HCV genotypes/subtypes were present. New variants of HCV genotype 6 were discovered in three samples from Kunming and one in Guangzhou in the Pearl River Delta.


Asunto(s)
Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C/epidemiología , China/epidemiología , ADN Viral/análisis , Variación Genética , Genotipo , Hepatitis C/virología , Humanos , Datos de Secuencia Molecular , Filogenia , ARN Viral/sangre , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Proteínas Virales/genética
12.
Intervirology ; 47(6): 289-309, 2004.
Artículo en Inglés | MEDLINE | ID: mdl-15564741

RESUMEN

Sequences of 234 complete genomes and 631 hepatitis B surface antigen genes were used to assess the worldwide diversity of hepatitis B virus (HBV). Apart from the described two subgenotypes each for A and F, also B, C, and D divided into four subgenotypes each in the analysis of complete genomes supported by significant bootstrap values. The subgenotypes of B and C differed in their geographical distribution, with B1 dominating in Japan, B2 in China and Vietnam, B3 confined to Indonesia, and B4 confined to Vietnam, all strains specifying subtype ayw1. Subgenotype C1 was common in Japan, Korea, and China; C2 in China, South-East Asia, and Bangladesh, and C3 in the Oceania comprising strains specifying adrq-, and C4 specifying ayw3 is encountered in Aborigines from Australia. This pattern of defined geographical distribution was less evident for D1-D4, where the subgenotypes were widely spread in Europe, Africa, and Asia, possibly due to their divergence having occurred a longer time ago than for genotypes B and C, with D4 being the first split and still the dominating subgenotype of D in the Oceania. The genetic diversity of HBV and the geographical distribution of its subgenotypes provide a tool to reconstruct the evolutionary history of HBV and may help to complement genetic data in the understanding of the evolution and past migrations of man.


Asunto(s)
Variación Genética/genética , Virus de la Hepatitis B/genética , Epidemiología Molecular , Secuencia de Aminoácidos , Animales , Demografía , Genotipo , Virus de la Hepatitis B/clasificación , Humanos , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia
13.
J Virol ; 78(22): 12252-8, 2004 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-15507612

RESUMEN

Acute hepatitis C virus (HCV) is typically defined as new viremia and antibody seroconversion. Rates and immunologic correlates of hepatitis C clearance have therefore been based on clearance of viremia only in individuals who initially had an antibody response. We sought to characterize the immunological correlates of clearance in patients with acute hepatitis C and their sexual contacts. We prospectively determined CD4(+) and CD8(+) cytotoxic T-lymphocyte responses in index patients with acute HCV and their sexual contacts who developed acute infection, either with or without spontaneous clearance, as well as those contacts who never developed viremia. Responses were measured using proliferation and ELISpot assays for CD4(+) and CD8(+) responses. We demonstrate in this prospective study that cellular immune responses can develop in exposed but persistently aviremic and antibody-negative individuals as well as those individuals with spontaneous clearance of acute HCV. These findings lend further credence to the importance of cellular immune responses in recovery from HCV and suggest that low exposure to HCV may lead to development of HCV-specific immune responses without ongoing HCV replication. This finding has important implications for HCV vaccine and therapeutic development.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/inmunología , Linfocitos T Citotóxicos/inmunología , Enfermedad Aguda , Adulto , Femenino , Hepatitis C/transmisión , Humanos , Interferón gamma/biosíntesis , Activación de Linfocitos , Masculino , Conducta Sexual
14.
J Med Virol ; 74(4): 556-62, 2004 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-15484284

RESUMEN

Hepatitis E virus (HEV) was identified by RT-PCR amplification with degenerate ORF2 primers in the stool of a piglet experimentally inoculated with a stool suspension from a patient with acute hepatitis during an outbreak of non-A, non-B hepatitis in Kyrgyzstan. Further characterization by sequencing of the complete genome and phylogenetic analysis showed that the piglet isolate was most closely related to HEV genotype 3. Because the original human stool specimen used to inoculate the piglet was no longer available, stool samples from three patients obtained during the same outbreak were sequenced and found to be HEV genotype 1. These findings suggest that the HEV isolated from the swine stool was probably an HEV enzootic in Kyrgyzstan and not the virus inoculated from the human stool.


Asunto(s)
Genoma Viral , Virus de la Hepatitis E/genética , Hepatitis E/veterinaria , Enfermedades de los Porcinos/virología , Animales , Cartilla de ADN/genética , Heces/virología , Hepatitis E/transmisión , Virus de la Hepatitis E/clasificación , Humanos , Kirguistán , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADN , Porcinos
15.
J Gen Virol ; 85(Pt 10): 2943-2952, 2004 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-15448357

RESUMEN

The complete genomic sequence of hepatitis A virus (HAV) CF53/Berne strain was determined. Pairwise comparison with other complete HAV genomic sequences demonstrated that the CF53/Berne isolate is most closely related to the single genotype VII strain, SLF88. This close relationship was confirmed by phylogenetic analyses of different genomic regions, and was most pronounced within the capsid region. These data indicated that CF53/Berne and SLF88 isolates are related more closely to each other than are subtypes IA and IB. A histogram of the genetic differences between HAV strains revealed four separate peaks. The distance values for CF53/Berne and SLF88 isolates fell within the peak that contained strains of the same subtype, showing that they should be subtypes within a single genotype. The complete genomic data indicated that genotypes II and VII should be considered a single genotype, based upon the complete VP1 sequence, and it is proposed that the CF53/Berne isolate be classified as genotype IIA and strain SLF88 as genotype IIB. The CF53/Berne isolate is cell-adapted, and therefore its sequence was compared to that of two other strains adapted to cell culture, HM-175/7 grown in MK-5 and GBM grown in FRhK-4 cells. Mutations found at nucleotides 3889, 4087 and 4222 that were associated with HAV attenuation and cell adaptation in HM175/7 and GMB strains were not present in the CF53/Berne strain. Deletions found in the 5'UTR and P3A regions of the CF53/Berne isolate that are common to cell-adapted HAV isolates were identified, however.


Asunto(s)
Genoma Viral , Virus de la Hepatitis A/genética , Secuencia de Bases , Genotipo , Virus de la Hepatitis A/clasificación , Datos de Secuencia Molecular , Filogenia
16.
J Med Virol ; 73(3): 378-83, 2004 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-15170631

RESUMEN

A limited number of hepatitis A virus (HAV) isolates from South America have been characterised at the genomic level. IgM anti-HAV positive serum samples collected from patients with hepatitis A living in the five geographical regions of Brazil (North, Northeast, Central, South, and Southeast) were used to obtain HAV isolates and determine their genetic relatedness. Of the 232 case isolates, sequence data were obtained from the VP1/2A junction region of the HAV genome. All isolates were classified in genotype I; 231 belonged to subgenotype IA, and one to subgenotype IB. HAV isolates from four States formed distinct clusters of highly related sequences. However, isolates from other states did not cluster and the sequences from those states were intermingled with sequences found in the other states. The amino acid sequences of all but two isolates showed a Leu --> Ile substitution at position 42 in the 2A protein. This substitution appeared to be a characteristic geographic fingerprint of HAV sequences within Brazil.


Asunto(s)
Genoma Viral , Virus de la Hepatitis A/genética , Virus de la Hepatitis A/aislamiento & purificación , Hepatitis A/virología , Adolescente , Adulto , Anciano , Sustitución de Aminoácidos , Brasil , Niño , Preescolar , Femenino , Genotipo , Virus de la Hepatitis A/clasificación , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , ARN Viral/sangre , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia , Proteínas Estructurales Virales/química , Proteínas Estructurales Virales/genética
17.
J Gen Virol ; 84(Pt 12): 3275-3289, 2003 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-14645909

RESUMEN

We determined the sequence of the hepatitis delta virus (HDV) genome in 40 Japanese patients, most of whom were from the Miyako Islands, Okinawa, Japan. Consensus sequences from 33 HDV full genomes out of a total of 40 patients were determined by directly sequencing four partially overlapping PCR products. Phylogenetic tree analysis classified these 33 complete HDV genomes as HDV genotype I (two patients), genotype IIa (one patient) and genotype IIb (30 patients). Among the 30 genotype IIb patients, there were two clusters of genetic variants. One group consisted of six isolates showing significant homology with genotype IIb, previously reported from Taiwan. The other group consisted of 24 isolates, whose sequences formed a new genetic subgroup (genotype IIb-Miyako; IIb-M). When the genetic structures were compared in detail between IIb and IIb-M, characteristic variations were found in the C-terminal sequence of the large delta antigen-conferring packaging signal as well as the RNA editing site. Determination of subclasses of genotype IIb in a total of 37 patients, including seven HDV patients whose partial HDV sequence was determined, revealed eight patients with IIb and 29 patients with IIb-M. Although there was no significant difference in the clinical background or virological state of hepatitis B virus between these two groups, patients with genotype IIb-M showed greater progression of chronic hepatitis and cirrhosis than those with genotype IIb (P=0.0009). These data indicate the existence of a genetic subgroup of HDV genotype IIb, which is associated with different clinical characteristics and which could be related to genetic variations in functionally important parts of the HDV genome.


Asunto(s)
Genoma Viral , Hepatitis D Crónica/diagnóstico , Virus de la Hepatitis Delta/genética , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Aminoácidos , Secuencia de Bases , Secuencia de Consenso , Femenino , Variación Genética , Genotipo , Hepatitis D Crónica/patología , Virus de la Hepatitis Delta/clasificación , Virus de la Hepatitis Delta/aislamiento & purificación , Antígenos de Hepatitis delta/genética , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia
18.
J Clin Microbiol ; 41(6): 2727-33, 2003 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-12791916

RESUMEN

The level of in vitro detection of viral genomes in mixes with two different hepatitis C virus (HCV) subtypes was investigated by artificially mixing previously measured subtype-specific HCV RNA genomes. The RNAs in these mixtures were reverse transcribed and then PCR amplified by using two sets of primers corresponding to the 5' untranslated region and digested with endonucleases to analyze the restriction fragment length polymorphism patterns. This approach facilitated detection of a wider range of type-specific HCV genomes than originally described, beyond equimolar concentrations of contributing HCV subtypes. Moreover, by using computerized image analysis, this study also demonstrated that the true contribution of each virus type-and consequently of mixed infections-may be underestimated when only visual observation is carried out. These results may be useful for comparing data obtained from this and other currently used methodologies.


Asunto(s)
Genoma Viral , Hepacivirus/clasificación , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/virología , Polimorfismo de Longitud del Fragmento de Restricción , ARN Viral/sangre , Genotipo , Hepacivirus/genética , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Especificidad de la Especie
19.
J Gen Virol ; 83(Pt 8): 2059-2073, 2002 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-12124470

RESUMEN

The complete genomes were sequenced for ten hepatitis B virus (HBV) strains. Two of them, from Spain and Sweden, were most similar to genotype D, although encoding d specificity. Five of them were from Central America and belonged to genotype F. Two strains from Nicaragua and one from Los Angeles, USA, showed divergences of 3.1-4.1% within the small S gene from genotype F strains and were recognized previously as a divergent clade within genotype F. The complete genomes of the two genotype D strains were found to differ from published genotype D strains by 2.8-4.6%. Their S genes encoded Lys(122), Thr(127) and Lys(160), corresponding to the putative new subtype adw3 within this genotype, previously known to specify ayw2, ayw3 or, rarely, ayw4. The complete genomes of the three divergent strains diverged by 0.8-2.5% from each other, 7.2-10.2% from genotype F strains and 13.2-15.7% from other HBV strains. Since pairwise comparisons of 82 complete HBV genomes of intratypic and intertypic divergences ranged from 0.1 to 7.4% and 6.8 to 17.1%, respectively, the three sequenced strains should represent a new HBV genotype, for which the designation H is proposed. In the polymerase region, the three strains had 16 unique conserved amino acid residues not present in genotype F strains. So far, genotype H has been encountered in Nicaragua, Mexico and California. Phylogenetic analysis of the complete genomes and subgenomes of the three strains showed them clustering with genotype F but forming a separate branch supported by 100% bootstrap. Being most similar to genotype F, known to be an Amerindian genotype, genotype H has most likely split off from genotype F within the New World.


Asunto(s)
Pueblo Asiatico , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B/etnología , Indígenas Norteamericanos , América Central/epidemiología , Variación Genética , Genoma Viral , Genotipo , Hepatitis B/epidemiología , Hepatitis B/virología , Antígenos de Superficie de la Hepatitis B/genética , Humanos , Datos de Secuencia Molecular , Nicaragua/epidemiología , Filogenia , Precursores de Proteínas/genética , Análisis de Secuencia de ADN
20.
Infect Control Hosp Epidemiol ; 23(6): 313-8, 2002 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-12083234

RESUMEN

OBJECTIVE: To identify exposures associated with acute hepatitis B virus (HBV) infection among residents with diabetes in a skilled nursing facility. DESIGN: Residents from Unit 3 and other skilled nursing facility residents with diabetes were tested for serologic evidence of HBV infection. Two retrospective cohort studies were conducted. Potential routes of HBV transmission were evaluated by statistical comparison of attack rates. SETTING: A 269-bed skilled nursing facility. PARTICIPANTS: All skilled nursing facility residents with diabetes and skilled nursing facility residents who lived on the same unit as the index case (Unit 3) for some time during the case's incubation period. RESULTS: All 5 residents with acute HBV infection had diabetes and resided in Unit 3. The attack rate among the 12 patients with diabetes in Unit 3 was 42%, compared with 0% among 43 patients without diabetes (relative risk, 37.2; 95% confidence interval, 4.7 to infinity). Acutely infected patients with diabetes received more morning insulin doses (P = .05), and more insulin doses (P = .03) and finger sticks (P = .02) on Wednesdays than did noninfected patients with diabetes. Two chronically infected patients with diabetes in Unit 3 were positive for hepatitis B e antigen and regularly received daily insulin and finger sticks. Of the 4 acute and 3 chronically infected residents from whom HBV DNA was amplified, all were genotype F and had an identical 678-bp S region sequence. Although no component of the lancets or injection devices was shared among residents, opportunities for HBV contamination of diabetes care supplies were identified. CONCLUSIONS: Contamination of diabetes care supplies resulted in resident-to-resident transmission of HBV. In any setting in which diabetes care is performed, staff need to be educated regarding appropriate infection control practices.


Asunto(s)
Infección Hospitalaria/transmisión , Diabetes Mellitus/sangre , Transmisión de Enfermedad Infecciosa , Hepatitis B/transmisión , Instituciones de Cuidados Especializados de Enfermería , Enfermedad Aguda , Anciano , Infección Hospitalaria/sangre , Complicaciones de la Diabetes , Femenino , Hepatitis B/sangre , Hepatitis B/complicaciones , Humanos , Masculino , Lesiones por Pinchazo de Aguja , Características de la Residencia , Estudios Retrospectivos
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