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By the end of 2021, the Omicron variant of SARS-CoV-2, the coronavirus responsible for COVID-19, emerges, causing immediate concern, due to the explosive increase in cases in South Africa and a large number of mutations. This study describes the characteristic mutations of the Omicron variant in the Spike protein, and the behavior of the successive epidemic waves associated to the sub-lineages throughout the world. The mutations in the Spike protein described are related to the virus ability to evade the protec-tion elicited by current vaccines, as well as with possible reduced susceptibil-ity to host proteases for priming of the fusion process, and how this might be related to changes in tropism, a replication enhanced in nasal epithelial cells, and reduced in pulmonary tissue; traits probably associated with the apparent reduced severity of Omicron compared to other variants.

Introducción: El agente etiológico responsable de COVID-19, SARSCoV-2, es un virus ARN perteneciente a la familia Coronaviridae. Durante la replicación, los componentes virales interactúan con la maquinaria celular induciendo alteraciones en la fisiología celular, lo que contribuye a la patogénesis del virus. Método: Revisión bibliográfica en NCBI/Pubmed sobre estrés celular y SARS-CoV-2 Hallazgos de interpretación: Como respuesta a la infección, en la célula hospedadora se activan vías de señalización, cuyo principal objetivo es recuperar la homeostasis y de no lograrlo, inducir a la activación de la muerte celular. Entre las vías de señalización mejor caracterizadas, destacan las rutas de estrés celular como el estrés oxidativo, la UPR (Respuesta a proteínas no plegadas), y la autofagia, las cuales son evolutivamente bien conservadas y además están interconectadas entre sí. Hay fuerte evidencia teórica y experimental de diversas interacciones de algunos componentes de estas rutas con distintas proteínas virales de los coronavirus, y ya se han adelantado algunos estudios con SARS-CoV-2. En esta revisión, resaltamos algunas de las rutas celulares-virus que se han caracterizado hasta el momento. Reflexiones finales: Aún queda mucho por entender de estas rutas y su relación con las infecciones virales; esto pudiera constituir un importante blanco para la investigación y desarrollo de terapias antivirales.

Introducción: La infección por SARS-CoV-2 determina desde formas asintomáticas hasta manifestaciones muy graves de COVID-19 que pueden comprometer la vida. El impacto de la pandemia por COVID-19 plantea la necesidad de identificar los factores de riesgo que condicionan la variabilidad interindividual en la susceptibilidad a la infección por SARS-CoV-2, la fisiopatología y la progresión de la enfermedad. Varios autores sugieren que el componente genético juega un papel clave en la compleja interacción existente virus-hospedero, donde las diferencias poblacionales dirigen tal heterogeneidad. Métodos: Se realizó una revisión bibliográfica de la literatura científica, publicada hasta los momentos, relacionada con los marcadores biológicos que participan en la evolución de la infección por SARS-CoV-2 y los polimorfismos de moléculas potencialmente involucradas en la compleja interacción virus-hospedero. Hallazgos de interpretación: Numerosas evidencias señalan que variantes alélicas de genes que codifican para moléculas que actúan en las etapas tempranas de interacción del virus con la célula o en eventos que determinan la respuesta inmunitaria del hospedero, podrían favorecer la infección por SARS-CoV-2 o la gravedad de la enfermedad. El grupo sanguíneo ABO pareciera jugar un papel en la inmunopatogénesis, mostrando los individuos de grupo O un menor riesgo a infectarse con SARS-CoV-2, aunque otros componentes del hospedero, genéticos, fisiológicos y metabólicos, como la expresión diferencial de receptores tipo toll, antígenos HLA, citocinas proinflamatorias, quimiocinas u otros efectores de la respuesta inmunitaria innata y dquirida, estarían también participando. Reflexiones finales: El éxito y progresión de la infección viral hacia el desarrollo de una manifestación clínica y su gravedad dependen, en gran medida, de la interacción entre factores virales y la respuesta del hospedero, que a su vez está condicionada por factores genéticos, epigenéticos y comorbilidades preexistentes. La contribución del componente genético plantea la potencial aplicación de estrategias preventivas dirigidas, la identificación de blancos terapéuticos y el desarrollo de nuevos fármacos.

Background Human cosavirus (HCoSV) is a new member of the Picornaviridae family, geographically widespread among humans. It has been suggested as a causative agent of acute gastroenteritis, but its pathogenicity is not currently certain. In HIV-infected subjects, diarrhoea is one of the most frequent gastrointestinal manifestations, whose aetiology remains often unexplained. Objectives To identify the cause of viral diarrhoea among HIV infected patients by molecular assays. Study design A total of 143 stool samples from HIV subjects with and without diarrhoea, were screened for conventional enteric viruses (rotavirus, adenovirus, norovirus and astrovirus) by molecular assays. The presence of HCoSV genome was investigated by nested RT-PCR for the 5’UTR region. Positive samples were further characterized by sequencing and phylogenetic analysis. Results Enteric viruses were more frequently found in diarrhoea cases (9/82) than controls (0/61) (p=0.007). HCoSV was detected in five (3.5%) of the subjects affected by diarrhoea. Phylogenetic analysis revealed the predominance of the HCoSV species D. One patient suffered a persistent cosavirus infection with a same strain and after eight months he had a fatal outcome. No other pathogens could be detected. Conclusions The results suggest a role of non-conventional enteric viruses, as HCoSV, as a potential opportunistic agent causing persistent infection and deterioration of the clinical conditions in HIV-infected patients. Screening procedures and monitoring including such viruses would be helpful in the clinical management of such patients.

In less than two years since SARS-CoV-2 emerged, the new coronavirus responsible for COVID-19, has accumulated a great number of mutations. Many of these mutations are located in the Spike protein and some of them confer to the virus higher transmissibility or partial resistance to antibody mediated neutralization. Viral variants with such confirmed abilities are designated by WHO as Variants of Concern (VOCs). The aim of this study was to monitor the introduction of variants and VOCs in Venezuela. A small fragment of the viral genome was sequenced for the detection of the most relevant mutations found in VOCs. This approach allowed the detection of Gamma VOC. Its presence was confirmed by complete genome sequencing. The Gamma VOC was detected in Venezuela since January 2021, and in March 2021 was predominant in the East and Central side of the country, representing more than 95% of cases sequenced in all the country in April-May 2021. In addition to the Gamma VOC, other isolates carrying the mutation E484K were also detected. The frequency of this mutation has been increasing worldwide, as shown in a survey of sequences carrying E484K mutation in GISAID, and was detected in Venezuela in many probable cases of reinfection. Complete genome sequencing of these cases allowed us to identify E484K mutation in association with Gamma VOC and other lineages. In conclusion, the strategy adopted in this study is suitable for genomic surveillance of variants for countries lacking robust genome sequencing capacities. In the period studied, Gamma VOC seems to have rapidly become the dominant variant throughout the country.

La glucosa-6-fosfato deshidrogenasa (G6PD) es una enzima citoplasmática que se encuentra distribuida en todas las células del organismo y que cataliza el primer paso de la vía de las pentosas en el cual la glucosa 6 fosfato (G6P) es convertida a 6-fosfogluconato (6FG) y el NADP reducido a NADPH, proceso indispensable para proteger a los eritrocitos del daño oxidativo. La deficiencia de la enzima G6PD es la eritroenzimopatía más común, es recesiva y ligada al cromosoma X, con amplia distribución mundial y elevada heterogeneidad genética y bioquímica. Se realizó una revisión sobre aspectos bioquímicos, estructura, genética, bases moleculares, defecto enzimático, prevalencia de la deficiencia en el mundo y en Venezuela, y el papel de la deficiencia de G6PD en el tratamiento de la malaria por el incremento en el riesgo de hemolisis que lleva consigo la tendencia de aumentar la dosis total de Primaquina en sujetos paludismo, particularmente en las regiones donde predomina la infección por Plasmodium vivax.

Porcine enteric sapovirus (PES) has been shown to cause diarrhea under experimental conditions in gnotobiotic piglets. However, the role of PES as enteric pathogens in porcine farms remains unclear. To further understand the PES-host interactions under field conditions, a serological survey was carried out. To this end the capsid gene of a PES isolate was cloned in the baculovirus expression system and an ELISA was developed based on virus-like particles from the baculovirus-expressed PES capsid protein. A total of 85 serum samples collected from pigs ranging from 8 weeks to over 54 weeks of age were analyzed. An overall seroprevalence to PESs of 62% was found, with significant differences (p0.05) found between ages. Pigs younger than 10 weeks old and older than 12 weeks old showed high seroprevalences (70-100%), while pigs aged 10-12 weeks showed no detectable serum antibodies levels. Our results suggest that PES infections are common in pigs and that passively acquired maternal antibodies are soon replaced by actively acquired antibodies, whose titers increase gradually with age and that probably are maintained during lifetime.

Background Glucose-6-phosphate dehydrogenase (G6PD) deficiency causes acute haemolytic anaemia triggered by oxidative drugs such as primaquine (PQ), used for Plasmodiumvivax malaria radical cure. However, in many endemic areas of vivax malaria, patients are treated with PQ without any evaluation of their G6PD status. Methods G6PD deficiency and its genetic heterogeneity were evaluated in northeastern and southeastern areas from Venezuela, Cajigal (Sucre state) and Sifontes (Bolívar state) municipalities, respectively. Blood samples from 664 randomly recruited unrelated individuals were screened for G6PD activity by a quantitative method. Mutation analysis for exons 4–8 of G6PD gen was performed on DNA isolated from G6PD-deficient (G6PDd) subjects through PCR–RFLP and direct DNA sequencing. Results Quantitative biochemical characterization revealed that overall 24 (3.6 %) subjects were G6PDd (average G6PD enzyme activity 4.5 ± 1.2 U/g Hb, moderately deficient, class III), while DNA analysis showed one or two mutated alleles in 19 of them (79.2 %). The G6PD A-202A/376G variant was the only detected in 17 (70.8 %) individuals, 13 of them hemizygous males and four heterozygous females. Two males carried only the 376A → G mutation. No other mutation was found in the analysed exons. Conclusions The G6PDd prevalence was as low as that one shown by nearby countries. This study contributes to the knowledge of the genetic background of Venezuelan population, especially of those living in malaria-endemic areas. Despite the high degree of genetic mixing described for Venezuelan population, a net predominance of the mild African G6PD A-202A/376G variant was observed among G6PDd subjects, suggesting a significant flow of G6PD genes from Africa to Americas, almost certainly introduced through African and/or Spanish immigrants during and after the colonization. The data suggest that 1:27 individuals of the studied population could be G6PDd and therefore at risk of haemolysis under precipitating factors. Information about PQ effect on G6PDd individuals carrying mild variant is limited, but since the regimen of 45 mg weekly dose for prevention of malaria relapse does not seem to be causing clinically significant haemolysis in people having the G6PD A-variant, a reasoned weighing of risk–benefit for its use in Venezuela should be done, when implementing public health strategies of control and elimination.

The immune response elicited by the rotavirus nonstructural protein NSP4 and its potential role in protection against rotavirus disease are not well understood. We investigated the serological response to NSP4 and its correlation with disease protection in sera from 110 children suffering acute diarrhea, associated or not with rotavirus, and from 26 children who were recipients of the rhesus rotavirus tetravalent (RRV-TV) vaccine. We used, as antigens in an enzyme-linked immunosorbent assay (ELISA), affinity-purified recombinant NSP4 (residues 85 to 175) from strains SA11, Wa, and RRV (genotypes A, B, and C, respectively) fused to glutathione S -transferase. Seroconversion to NSP4 was observed in 54% (42/78) of the children who suffered from natural rotavirus infection and in 8% (2/26) of the RRV-TV vaccine recipients. Our findings indicate that NSP4 evokes significantly ( P < 0.05) higher seroconversion rates after natural infection than after RRV-TV vaccination. The serum antibody levels to NSP4 were modest (titers of ≤200) in most of the infected and vaccinated children. A heterotypic NSP4 response was detected in 48% of the naturally rotavirus-infected children with a detectable response to NSP4. Following natural infection or RRV-TV vaccination, NSP4 was significantly less immunogenic than the VP6 protein when these responses were independently measured by ELISA. A significant ( P < 0.05) proportion of children who did not develop diarrhea associated with rotavirus had antibodies to NSP4 in acute-phase serum, suggesting that serum antibodies against NSP4 might correlate with protection from rotavirus diarrhea. In addition, previous exposures to rotavirus did not affect the NSP4 seroconversion rate.

Several studies have shown that adaptation of various viruses to grow in certain cell lines of vertebrates, leads to the selection of virus variants that bind heparan sulfate (HS) with high affinity. In this study we investigated the susceptibility of strains of dengue virus (DENV) to oversulfated heparin an analogue of HS after passages in BHK-21 cells. Field isolates of the four serotypes of DENV with a limited number of passes in mosquito cells C6/36HT were serially passaged eight times in BHK-21 cells. The adaptation of the DENV to the cell culture selected viral variants with an increased replicative capacity in BHK-21 cells and an increased susceptibility to heparin compared with the original not adapted strains, with a more significant inhibition of the infectivity in DENV-2 and DENV-4.The E protein of the adapted strains showed changes in the amino acid sequence, particularly at the position K204R to DENV-1, N67K to DENV-2, K308R and V452A for DENV-3 and E327G to DENV-4. These substitutions implicated a gain of basic residues that increased the net positive charge of the protein. These results suggest that adaptation of DENV strains to BHK-21 cells implies changes in the envelope protein, changes associated to the protein reactivity with heparin, the inhibitory effectiveness of this compound varying depending on the viral strain. In addition, these results suggest that the HS can play an important role in the infectivity of the DENV strains adapted to vertebrate cell culture, but not in the infectivity of non-adapted DENV isolates.

108 rotavirus strains obtained from children with diarrhea hospitalized in Palermo, Italy, in the years 1990-1994, were examined by seminested PCR to study the relative frequency and distribution of the four most common alleles of the gene 4. Such strains were selected from 344 human rotavirus strains recovered in palermo during those years after characterization by electropherotyping, subgrouping and G serotyping. One hundred and seven of the 108 strains could be classified into P types, the P[8], G1 (38.3%) and the P[8], G4 (52.3%) types being predominant. The unique strain whose P genotype could not be identified showed an unusual combination of long migration electrophoretic pattern and subgroup I specificity.

Gastrointestinal disorders or GID are debilitating conditions common in individuals infected by the human immunodeficiency virus (HIV), capable of leading to death. Numerous etiological agents and pathophysiological mechanisms have been involved in this status. Although the use of highly active antiretroviral therapy (HAART) in many countries has greatly reduced the prevalence of gastrointestinal infections, enteric pathogens such as bacteria, parasites, fungi and viruses may still act as opportunist agents in these patients. Cytomegalovirus, adenovirus, calicivirus, astrovirus, rotavirus, enterovirus, picobirnavirus and some more recently described, like bocavirus and Aichi virus, have been detected in HIV patients. However, except for cytomegalovirus, which is an established etiological agent of GID in these patients, the role of the other viruses remains unclear. Several species of Cryptosporidium, microsporidia, Salmonella, atipical mycobacteria and Campylobacter jejuni, have also been recognized as important causes of GID in HIV patients. The progressive incorporation of increasingly sensitive immunological and molecular assays for antigen, antibody and pathogens detection from faeces, has improved the diagnosis of diarrhea and contributed to clarify the etiological significance of some microorganisms in immunocompetent patients. In Venezuela, some information is available about the prevalence of enteric pathogens in immunocompromised patients infected with HIV. The identification of the etiologic agent responsible for this condition may be useful for the management and treatment of these patients, for whom viral enteritis is a disease, which reduces their quality of life and causes a high public health spending.

To examine the role of enteric adenoviruses (Ad40 and Ad41) in children with acute gastroenteritis, we evaluated 273 children with diarrhoea and 137 without enteric symptoms in Palermo, Italy, during an 8-month period. Stools were tested by two home-made monoclonal-based ELISAs to detected genus-specific adenovirus antigen and to type Ad40 and Ad41. Twenty-five samples (6.1%) were found to contain adenovirus, 18 of which were grown in Graham 293 and in HEp-2 cells. Ad40 and Ad41 were detected in 2.6% of children with diarrhoea and in none in the control group, while non-enteric adenoviruses were obtained from both patients (3.2%) and controls (6.5%). Samples containing Ad40 and Ad41 were positive by the virus isolation procedure in Graham and in HEp-2 cells, showing no distinct growth pattern in these cell lines. The evaluation of a latex agglutination test (Adenolex) and of a commercial ELISA (Adenoclone), respectively available for the detection of genus adenovirus antigen and for the typing of Ad40 and Ad41 suggests that both tests enable the identification of enteric adenoviruses in stool specimens, giving results comparable to our ELISAs.

Rotavirus infection induces an increase in [Ca2+]cyto, which in turn may affect the distribution of the cytoskeleton proteins in the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5 h.p.i. using fluorescent phalloidin. Western blot analysis indicated that RhoA is activated between 0.5 and 1 h.p.i. Neither the phosphorylation of RhoA nor the formation of stress fibers were observed in cells infected with virions pre-treated with an anti-VP5* non-neutralizing mAb, suggesting that RhoA activation is stimulated by the interaction of the virus with integrins forming the cell receptor complex. In addition, the structure of the tubulin cytoskeleton was also studied. Alterations of the microtubules were evident starting at 3 h.p.i. and by 7 h.p.i. when microtubules were markedly displaced toward the periphery of the cell cytoplasm. Loading of rotavirus-infected cells with either a Ca2+ chelator (BAPTA) or transfection with siRNAs to silence NSP4, reversed the changes observed in both the microfilaments and microtubules distribution, but not the appearance of stress fibers. These results indicate that alterations in the distribution of actin microfilaments are initiated early during infection by the activation of RhoA, and that latter changes in the Ca2+ homeostasis promoted by NSP4 during infection may be responsible for other alterations in the actin and tubulin cytoskeleton. publishedVersion

Rotavirus infection is the most common cause of severe gastroenteritis during childhood worldwide, especially in developing countries. Two rotavirus vaccines are available for childhood immunization programs. Evaluation of the vaccine performance will benefit from knowledge of the epidemiological features of rotavirus infection in regional settings. Limited information on the molecular characteristics of the rotavirus types circulating in Venezuela is available. Eighty seven (89.7%) of the 97 ELISA rotavirus positive stool samples collected from children with diarrhea aged

The circulation of Aichi virus in a major urban area was demonstrated using molecular detection with samples recovered from a major river polluted with sewage discharges in Caracas, Venezuela. Five out of 11 water samples studied were positive, being classified by phylogenetic analysis as genotype B. Analysis of sewage waters appears to be a useful methodology to uncover the presence of a hitherto undetected fecal pathogen in a given geographical area.

The detection and molecular characterization of pathogenic human viruses in urban sewage have been used extensively to derive information on circulating viruses in given populations throughout the world. In this study, a similar approach was applied to provide an overview of the epidemiology of waterborne gastroenteritis viruses circulating in urban areas of Caracas, the capital city of Venezuela in South America. Dry season sampling was conducted in sewers and in a major river severely polluted with urban sewage discharges. Nested PCR was used for detection of human adenoviruses (HAds), while reverse transcription plus nested or seminested PCR was used for detection of enteroviruses (HuEVs), rotaviruses (HRVs), noroviruses (HuNoVs), and astroviruses (HAstVs). HRVs were fully characterized with genotype-specific primers for VP4 (genotype P), VP7 (genotype G), and the rotavirus nonstructural protein 4 (NSP4). HuNoVs and HAstVs were characterized by sequencing and phylogenetic analysis. The detection rates of all viruses were ≥50%, and all sampling events were positive for at least one of the pathogenic viruses studied. The predominant HRV types found were G1, P[8], P[4], and NSP4A and -B. Genogroup II of HuNoVs and HAstV type 8 were frequently detected in sewage and sewage-polluted river waters. This study reveals relevant epidemiological data on the distribution and persistence of human pathogenic viruses in sewage-polluted waters and addresses the potential health risks associated with transmission of these viruses through water-related environmental routes.

Rotavirus infection of cells in culture induces major changes in Ca 2+ homeostasis. These changes include increases in plasma membrane Ca 2+ permeability, cytosolic Ca 2+ concentration, and total cell Ca 2+ content and a reduction in the amount of Ca 2+ released from intracellular pools sensitive to agonists. Various lines of evidence suggest that the nonstructural glycoprotein NSP4 and possibly the major outer capsid glycoprotein VP7 are responsible for these effects. In order to evaluate the functional roles of NSP4 and other rotavirus proteins in the changes in Ca 2+ homeostasis observed in infected cells, the expressions of NSP4, VP7, and VP4 were silenced using the short interfering RNA (siRNA) technique. The transfection of specific siRNAs resulted in a strong and specific reduction of the expression of NSP4, VP7, and VP4 and decreased the yield of new viral progeny by more than 90%. Using fura-2 loaded cells, we observed that knocking down the expression of NSP4 totally prevented the increase in Ca 2+ permeability of the plasma membrane and cytosolic Ca 2+ concentration measured in infected cells. A reduction in the levels of VP7 expression partially reduced the effect of infection on plasma membrane Ca 2+ permeability and Ca 2+ pools released by agonist (ATP). In addition, the increase of total Ca 2+ content (as measured by 45 Ca 2+ uptake) observed in infected cells was reduced to the levels in mock-infected cells when NSP4 and VP7 were silenced. Finally, when the expression of VP4 was silenced, none of the disturbances of Ca 2+ homeostasis caused by rotaviruses in infected cells were affected. These data altogether indicate that NSP4 is the main protein responsible for the changes in Ca 2+ homeostasis observed in rotavirus-infected cultured cells. Nevertheless, VP7 may contribute to these effects.

A case of chronic intestinal infection due to adenovirus type 40 lasting for 13 months in a patient with AIDS is described. Adenovirus particles were detected by electron microscopy in biopsy samples taken from the duodenum 3 months after the onset of diarrhoea. The virus was identified as adenovirus type 40 in stool samples by ELISA monoclonal antibodies to adenovirus group antigen (MAd-g2) and types 40 and 41 (MA 40-1 and MA 41-1). No other enteropathogens were found. These data support a causal relationship between adenovirus 40 and the gastrointestinal symptoms of the patient. This is the first reported case of intestinal infection caused by adenovirus type 40 in a patient with AIDS.

Polyacrylamide gel electrophoresis of nucleic acid extracted from stool samples of diarrhoeic children revealed in 3 out of 690 (0.43 %) specimens two electrophoretic bands with a migration pattern characteristic of picobirnavirus ds-RNA. In none of the 92 control children were similar bands detected. No other potential enteric pathogens were found in the patients with picobirnavirus infection.

The importance of enteric viral infections in HIV-related diarrhea is uncertain. Human caliciviruses have emerged as a leading cause of acute diarrhea worldwide.To evaluate the importance of calicivirus infections in HIV-related diarrhea. Study design 151 fecal samples collected from children and adults infected with HIV, with and without diarrhea, were examined. In addition, 89 fecal samples from non HIV-infected children and adults were also tested. Samples were analyzed by RT-PCR using primer sets specific to Norovirus genogroup I or genogroup II as well as primers designed to react with both Noroviruses and Sapovirus genus.Viruses were detected with equal frequencies in stools from HIV infected and non-infected adults (12%). However, specimens from HIV infected children were more likely than those of HIV-negative children to have caliciviruses (51% versus 24%, P0.05). Viral infections were not significantly associated with diarrhea neither in children nor in adults, regardless of HIV status. Viruses genetically related to the common Lordsdale virus (Norovirus genogroup II) and London/92 virus (Sapovirus) clusters were detected circulating among children.These results suggest that caliciviruses may be an important opportunistic pathogen in children infected with HIV.

Caliciviruses are a well-established cause of respiratory, vesicular and hemorrhagic diseases in animals. In addition, these viruses are an important cause of enteric diseases in humans. Recently, molecular analysis of several bovine enteric calicivirus isolates indicated that they are genetically close to human enteric calicivirus. To investigate if bovine enteric caliciviruses circulate in Venezuela, 129 stool samples collected between 1994 and 2000 were assayed by reverse transcription-polymerase chain reaction amplification. The presence of calicivirus was confirmed in one of the samples analyzed, collected in the Lara State from a healthy calf, 2 months old. Phylogenetic studies based on partial RNA polymerase sequences indicated that the Venezuelan isolate (Bo/NV/Lara/2000/VE) is most closely related to the genogroup III, genus Noroviruses.

We report a significant high incidence of infection with G9P[8] rotavirus in Italian children during the winter epidemic season 1999-2000. The study was carried out on 439 children < 4 years hospitalized with acute diarrhea in Palermo. G9P[8] strains constituted 19% of all rotavirus identified and were not associated with more severe forms of gastroenteritis.

The aim of our study was to determine whether the severity of rotavirus gastroenteritis may be related to the different characteristics of infecting viral strains. The severity of clinical symptoms in 401 children with acute rotavirus gastroenteritis was assessed using a scoring system for frequency and duration of vomiting, diarrhea, and fever, as well as the patients' requirements for intravenous rehydration. Rotavirus strains were characterized by determining the electropherotype of their double-stranded RNA, the G type and subgroup by a panel of monoclonal antibodies, and the P type by reverse transcription-polymerase chain reaction. Strains with a short electropherotype, G2P[4] type, and subgroup I were associated with more-severe gastroenteritis and affected children older than those infected with strains with a long electropherotype, G1P[8] or G4P[8] type, and subgroup II. Minor differences in clinical symptoms were also detected in children infected with different long electropherotypes and with G1P[8] and G4P[8] specificities.

A seminested PCR typing assay has been extended to identify rotavirus strains with the P[14] genotype. The specificity of the method was confirmed by Southern hybridization and by restriction analysis with the enzyme Alu I. One out of four human rotavirus (HRV) strains with unusual subgroup-electropherotype linkage but none out of 50 HRV strains with usual linkage was typed as P[14].

Summary During a measles outbreak, 112 serum specimens from 88 hospitalized patients were received in our laboratory for investigation of a morbilliform rash. These specimens (88 acute- and 24 convalescent-phase) were tested for the presence of measles-specific IgM antibodies by a capture EIA (enzyme immunoassay) using peroxidase-conjugated measles virus antigens and by an indirect EIA. Commercially available indirect EIA kits for measles-specific IgM antibodies were also used and compared with our homemade EIAs. Specificity studies included a collection of serum specimens containing rheumatoid factor, antinuclear antibodies or IgM antibodies specific to other viruses, and sera from blood donors and healthy children. Sensitivity of capture EIA and indirect EIA to detect measles IgM was 91.8 and 90.3%, respectively, and specificity was 98.2% for both tests. Specific IgM antibodies were detected in 70.5% of serum specimens at the first day after rash onset and were present for a month following the rash. Among the commercial measles IgM detection assays, EIA “Behring” was found to be a valid alternative for detection of measles virus-specific IgM.

Background Rotavirus (RV) is the most common cause of severe childhood diarrhea worldwide. Despite Venezuela was among the first developing countries to introduce RV vaccines into their national immunization schedules, RV is still contributing to the burden of diarrhea. Concerns exist about the selective pressure that RV vaccines could exert on the predominant types and/or emergence of new strains. Results To assess the impact of RV vaccines on the genotype distribution 1 year after the vaccination was implemented, a total of 912 fecal specimens, collected from children with acute gastroenteritis in Caracas from February 2007 to April 2008, were screened, of which 169 (18.5%) were confirmed to be RV positive by PAGE. Rotavirus-associated diarrhea occurred all year-round, although prevailed during the coolest and driest months among unvaccinated children under 24 months old. Of 165 RV strains genotyped for G (VP7) and P (VP4) by seminested multiplex RT-PCR, 77 (46.7%) were G2P[4] and 63 (38.2%) G1P[8]. G9P[8], G3P[8] and G2P[6] were found in a lower proportion (7.3%). Remarkable was also the detection of