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1.
Exp Appl Acarol ; 88(1): 41-55, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36190668

RESUMO

House dust mite is a common cause of atopic dermatitis (AD) both in humans and dogs. Detection of serum IgE to allergens is commonly used to diagnose allergic diseases. However, false-positive reactions due to cross-reactivity and non-specific reactivity may lead to misdiagnosis. We compared human and canine IgE reactivities to mite component allergens. Canine IgE-reactive components of Dermatophagoides farinae and Tyrophagus putrescentiae were identified by tandem mass spectrometry. Recombinant proteins were produced and IgE reactivities to component allergens were assessed by ELISA and inhibition assays using sera from AD patients and dogs. Canine IgE-reactive proteins (Der f 1, Der f 11, Tyr p 4, Tyr p 8, Tyr p 11, Tyr p 28) were identified by proteome analysis. Most patients were sensitized to Der f 1 (93.3%) and Der f 2 (86.7%). Dogs showed high sensitization to Der f 2 (94.1%) and Der f 18 (84.6%). Both patients and dogs showed low IgE binding frequency to Tyr p 8, 43.3% and 4%, respectively. The ELISA inhibition study indicated that canine IgE reactivity to T. putrescentiae is mostly due to non-specific reaction and cross-reaction with D. farinae. Different IgE sensitization patterns were shown between allergic humans and dogs with AD, especially to Der f 18, for the first time in Korea. Furthermore, non-specific canine IgE reactivity to storage mite indicates the possibility of misdiagnoses. Standardizations focused on the major canine allergen content of extracts should be developed. This will allow precision diagnosis and individuated treatments for each patient and atopic dog.


Assuntos
Acaridae , Dermatite Atópica , Doenças do Cão , Hipersensibilidade , Humanos , Cães , Animais , Acaridae/metabolismo , Dermatite Atópica/diagnóstico , Dermatite Atópica/veterinária , Imunoglobulina E , Antígenos de Dermatophagoides , Pyroglyphidae , Alérgenos/análise , Poeira , Doenças do Cão/diagnóstico
2.
BMC Vet Res ; 17(1): 367, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34852811

RESUMO

BACKGROUND: Diarrhea is one of the most common clinical symptoms in cats and can be caused by infectious pathogens and investigation of the prevalence, co-infection and seasonality of enteropathogens are not well-established in diarrheic cats. RESULTS: Fecal samples of 1620 diarrheic cats were collected and enteropathogens were detected using real-time PCR. We retrospectively investigated the clinical features, total/seasonal prevalence, and infection patterns of enteropathogens. The positive infection rate was 82.59%. Bacterial, viral, and protozoal infections accounted for 49.3, 37.57, and 13.13% of cases, respectively. Feline enteric coronavirus (FECV) was the most common pathogen (29.37%), followed by Clostridium (C.) perfringens, Campylobacter (C.) coli, feline parvovirus, and Tritrichomonas foetus. The seasonality of enteropathogens was observed with peaks as follows: bacterial infections peaked in October, viral infections peaked in November, and protozoal infections peaked in August. Viral and protozoal infections showed differences in prevalence according to patient age. In the infection patterns, the ratios of single infections, mixed infections, and co-infections were 35.72, 9.87, and 54.41%, respectively. FECV was predominant in single infections. The most common patterns of multiple infections were C. perfringens and C. coli in mixed infections and C. perfringens and FECV in co-infections. CONCLUSIONS: Infection patterns differed according to the enteropathogen species, seasonality, and age distribution in cats. The results of this study might be helpful to understand in clinical characteristics of feline infectious diarrhea. In addition, continued monitoring of feline enteropathogens is required.


Assuntos
Doenças do Gato/virologia , Infecções por Coronavirus/veterinária , Coronavirus Felino/isolamento & purificação , Diarreia/veterinária , Fezes/microbiologia , Vírus da Panleucopenia Felina/isolamento & purificação , Animais , Campylobacter , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/veterinária , Gatos , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Infecções por Clostridium/veterinária , Clostridium perfringens , Coinfecção/veterinária , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Diarreia/virologia , Panleucopenia Felina/epidemiologia , Panleucopenia Felina/virologia , Prevalência , República da Coreia , Estudos Retrospectivos
3.
Korean J Parasitol ; 58(3): 257-265, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32615739

RESUMO

The outbreak of human toxoplasmosis can be attributed to ingestion of food contaminated with Toxoplasma gondii. Toxoplasmosis recently increased in domestic and stray dogs and cats. It prompted studies on the zoonotic infectious diseases transmitted via these animals. Sero- and antigen prevalences of T. gondii in dogs and cats were surveyed using ELISA and PCR, and B1 gene phylogeny was analyzed in this study. Toxoplasmosis antibodies were measured on sera of 403 stray cats, 947 stray dogs, 909 domestic cats, and 2,412 domestic dogs collected at nationwide regions, Korea from 2017 to 2019. In addition, whole blood, feces, and tissue samples were also collected from stray cats (1,392), stray dogs (686), domestic cats (3,040), and domestic dogs (1,974), and T. gondii-specific B1 gene PCR was performed. Antibody prevalence of stray cats, stray dogs, domestic cats, and domestic dogs were 14.1%, 5.6%, 2.3%, and 0.04%, respectively. Antigen prevalence of these animals was 0.5%, 0.2%, 0.1%, and 0.4%, respectively. Stray cats revealed the highest infection rate of toxoplasmosis, followed by stray dogs, domestic cats, and domestic dogs. B1 gene positives were 5 of stray cats, and identified to high/moderate pathogenic Type I/III group. These findings enforce that preventive hygienic measure should be strengthened at One Health level in dogs and cats, domestic and stray, to minimize human toxoplasmosis infections.


Assuntos
Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães/parasitologia , Genes de Protozoários/genética , Toxoplasma/genética , Toxoplasmose Animal/parasitologia , Animais , Filogenia , Reação em Cadeia da Polimerase , República da Coreia/epidemiologia , Estudos Soroepidemiológicos , Toxoplasmose/parasitologia , Toxoplasmose/prevenção & controle
4.
Korean J Parasitol ; 56(4): 371-374, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30196670

RESUMO

A 3-month-old female Maltese puppy was hospitalized with persistent diarrhea in a local veterinary clinic. Blood chemistry and hematology profile were analyzed and fecal smear was examined. Diarrheal stools were examined in a diagnostic laboratory, using multiplex real-time polymerase chain reaction (PCR) against 23 diarrheal pathogens. Sequence analysis was performed using nested PCR amplicon of 18S ribosomal RNA. Coccidian oocysts were identified in the fecal smear. Although multiplex real-time PCR was positive for Cyclospora cayetanensis, the final diagnosis was Cystoisospora ohioensis infection, confirmed by phylogenetic analysis of 18S rRNA. To our knowledge, this the first case report of C. ohioensis in Korea, using microscopic examination and phylogenetic analysis.


Assuntos
Coccídios/isolamento & purificação , Coccidiose/parasitologia , Coccidiose/veterinária , Diarreia/parasitologia , Diarreia/veterinária , Doenças do Cão/parasitologia , Enteropatias Parasitárias/parasitologia , Enteropatias Parasitárias/veterinária , Oocistos/isolamento & purificação , Animais , Coccídios/genética , Cães , Fezes/parasitologia , Feminino , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 18S , República da Coreia
5.
Virol J ; 12: 27, 2015 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-25886579

RESUMO

BACKGROUND: Enteroviruses (EVs) are the leading cause of aseptic meningitis worldwide. Detection of enteroviral RNA in clinical specimens has been demonstrated to improve the management of patient care, especially that of neonates and young children. FINDINGS: To establish a sensitive and reliable assay for routine laboratory diagnosis, we compared the sensitivity and specificity of the GeneXpert Enterovirus Assay (GXEA) with that of the reverse transcription polymerase chain reaction (RT-PCR) based assay referred to as real-time one step RT-PCR (RTo-PCR). The sensitivity/specificity produced by GXEA and RTo-PCR were 100%/100% and 65%/100%, respectively. CONCLUSIONS: Both methods evaluated in this article can be used for detection of enterovirus in clinical specimens and these nucleic acid amplification methods are useful assays for the diagnosis of enteroviral infection.


Assuntos
Infecções por Enterovirus/virologia , Enterovirus/isolamento & purificação , Meningite/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Enterovirus/genética , Infecções por Enterovirus/líquido cefalorraquidiano , Infecções por Enterovirus/diagnóstico , Humanos , Meningite/líquido cefalorraquidiano , Meningite/diagnóstico , Técnicas de Diagnóstico Molecular , RNA Viral/líquido cefalorraquidiano
6.
J Korean Med Sci ; 30(10): 1471-5, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26425045

RESUMO

Rotavirus (RV) is one of the most important viral etiologic agents of acute gastroenteritis (AGE) in children. Although effective RV vaccines (RVVs) are now used worldwide, novel genotypes and outbreaks resulting from rare genotype combinations have emerged. This study documented RV genotypes in a Korean population of children with AGE 5 yr after the introduction of RVV and assessed potential genotype differences based on vaccination status or vaccine type. Children less than 5-yr-old diagnosed with AGE between October 2012 and September 2013 admitted to 9 medical institutions from 8 provinces in Korea were prospectively enrolled. Stool samples were tested for RV by enzyme immunoassay and genotyped by multiplex reverse-transcription polymerase chain reaction. In 346 patients, 114 (32.9%) were RV-positive. Among them, 87 (76.3%) patients were infected with RV alone. Eighty-six of 114 RV-positive stool samples were successfully genotyped, and their combinations of genotypes were G1P[8] (36, 41.9%), G2P[4] (12, 14.0%), and G3P[8] (6, 7.0%). RV was detected in 27.8% of patients in the vaccinated group and 39.8% in the unvaccinated group (P=0.035). Vaccination history was available for 67 of 86 cases with successfully genotyped RV-positive stool samples; RotaTeq (20, 29.9%), Rotarix (7, 10.4%), unvaccinated (40, 59.7%). The incidence of RV AGE is lower in the RV-vaccinated group compared to the unvaccinated group with no evidence of substitution with unusual genotype combinations.


Assuntos
Vacinação em Massa , Infecções por Rotavirus/prevenção & controle , Vacinas contra Rotavirus/imunologia , Rotavirus/classificação , Rotavirus/genética , Pré-Escolar , Fezes/virologia , Gastroenterite/imunologia , Gastroenterite/prevenção & controle , Gastroenterite/virologia , Genótipo , Humanos , Lactente , RNA Viral/genética , República da Coreia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/isolamento & purificação , Infecções por Rotavirus/imunologia , Infecções por Rotavirus/virologia , Vacinas Atenuadas/imunologia
7.
J Med Virol ; 86(9): 1584-9, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24114945

RESUMO

Acute flaccid paralysis (AFP) is described as sudden onset of flaccid paralysis in one or more limbs in children caused by polioviruses (PVs). PV eradication is achieved through intensive immunization and AFP attentive surveillance, according to the World Health Organization. Since 1998, the Korea Centers for Disease Control and Prevention has conducted surveillance system. This is an overview of surveillance in the Republic of Korea during the 10-year period from 2002 to 2011. The surveillance system for wild PV eradication was conducted through reporting and laboratory testing. Cell culture isolates were identified by neutralization tests using standard polyclonal antisera typing. The molecular methods were used for further characterization to improve specificity. For genotyping, semi-nested RT-PCR was used to amplify part of the viral protein 1 gene. Patients below 5 years of age accounted for the largest proportion of cases, and a positive association between age and incidence was found. In the total 285 cases, Guillain-Barré syndrome was the major leading causes of AFP. Non-polio enterovirus was detected in some AFP patients. EV71 was detected in 21 cases and Coxsackievirus (C) A2, CA6, CA9, CB2, CB3, CB4, CB5, and Echovirus (E) 25, E30, Sabin strain polio 2, polio 1 and 3 were also detected in some patients. The present study represents a comprehensive 10-year country-based survey of AFP in the Republic of Korea. This surveillance could provide better understanding of the epidemiologic pattern, and clinical manifestations associated with specific genotypes of AFP in the Republic of Korea.


Assuntos
Enterovirus Humano A , Infecções por Enterovirus/epidemiologia , Síndrome de Guillain-Barré/epidemiologia , Paralisia/epidemiologia , Doença Aguda , Adolescente , Criança , Pré-Escolar , Surtos de Doenças , Feminino , Síndrome de Guillain-Barré/virologia , Humanos , Lactente , Recém-Nascido , Masculino , Paralisia/virologia , República da Coreia/epidemiologia
8.
Emerg Infect Dis ; 19(8): 1268-75, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23876671

RESUMO

The epidemiology of enteroviral infection in South Korea during 1999-2011 chronicles nationwide outbreaks and changing detection and subtyping methods used over the 13-year period. Of 14,657 patients whose samples were tested, 4,762 (32.5%) samples were positive for human enterovirus (human EV); as diagnostic methods improved, the rate of positive results increased. A seasonal trend of outbreaks was documented. Genotypes enterovirus 71, echovirus 30, coxsackievirus B5, enterovirus 6, and coxsackievirus B2 were the most common genotypes identified. Accurate test results correlated clinical syndromes to enterovirus genotypes: aseptic meningitis to echovirus 30, enterovirus 6, and coxsackievirus B5; hand, foot and mouth disease to coxsackievirus A16; and hand, foot and mouth disease with neurologic complications to enterovirus 71. There are currently no treatments specific to human EV infections; surveillance of enterovirus infections such as this study provides may assist with evaluating the need to research and develop treatments for infections caused by virulent human EV genotypes.


Assuntos
Surtos de Doenças , Infecções por Enterovirus/diagnóstico , Enterovirus/genética , Adolescente , Adulto , Distribuição por Idade , Proteínas do Capsídeo/genética , Criança , Pré-Escolar , Erros de Diagnóstico , Infecções por Enterovirus/epidemiologia , Monitoramento Epidemiológico , Genótipo , Humanos , Lactente , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular , Tipagem Molecular , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , República da Coreia/epidemiologia , Estações do Ano , Sensibilidade e Especificidade , Análise de Sequência de DNA , Adulto Jovem
9.
J Clin Microbiol ; 51(2): 598-600, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23175251

RESUMO

Prevalence of asymptomatic norovirus infection was investigated in food handlers in South Korea. Among 6,441 subjects, 66 (1.02%) had norovirus infections confirmed by reverse transcription (RT)-PCR (real time and nested). GII-12 and GII-4 were the prevalent genotypes. Our data suggest that infection of asymptomatic food handlers is an important transmission source in norovirus outbreaks.


Assuntos
Infecções por Caliciviridae/epidemiologia , Manipulação de Alimentos , Norovirus/genética , Infecções por Caliciviridae/transmissão , Proteínas do Capsídeo/genética , Surtos de Doenças , Genótipo , Humanos , Dados de Sequência Molecular , Norovirus/classificação , Filogenia , Prevalência , República da Coreia/epidemiologia , Estações do Ano
10.
J Med Virol ; 85(7): 1274-9, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23918544

RESUMO

Human enterovirus (EV) 71 is the main etiological agent of hand, foot, and mouth disease (HFMD). It is associated with neurological complications, and caused fatalities during recent outbreaks in the Asia-Pacific region. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. In this study, a duplex real-time RT-PCR assay was developed in order to simultaneously detect pan-EV and EV71. EV71-specific primers and probes were designed based on the highly conserved VP1 region of EV71. Five EV71 strains were detected as positive, and no positive fluorescence signal was observed in the duplex real-time RT-PCR for other viral RNA, which showed 100% specificity for the selected panel, and no cross-reactions were observed in this duplex real-time RT-PCR. The EV71-specific duplex real-time RT-PCR was more sensitive than conventional RT-PCR, and detected viral titers that were 10-fold lower than those measured by the latter. Of the 381 HFMD clinical specimens, 196 (51.4%) cases were pan-EV-positive, of which 170 (86.7%) were EV71-positive when tested by pan-EV and EV71-specific duplex real-time RT-PCR. EV71-specific duplex real-time RT-PCR offers a rapid and sensitive method to detect EV71 from clinical specimens, and will allow quarantine measures to be taken more effectively during outbreaks.


Assuntos
Enterovirus Humano A/isolamento & purificação , Doença de Mão, Pé e Boca/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Virologia/métodos , Animais , Criança , Pré-Escolar , Enterovirus Humano A/genética , Doença de Mão, Pé e Boca/virologia , Humanos , Sensibilidade e Especificidade , Fatores de Tempo
11.
J Med Virol ; 85(1): 110-5, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23073968

RESUMO

This study was performed to analyze epidemiological and molecular characteristics of coxsakievirus (CV) B1 infection associated with severe neonatal illness cases and death in Korea during 2008-2009. Through a nationwide surveillance program, specimens were collected from 104 patients infected with CVB1. The detection of enteroviruses (EVs) from specimens was subjected to a diagnostic real-time polymerase chain reaction (RT-PCR) in the 5'-non-coding region (NCR). A semi-nested PCR was conducted to amplify sequences from the VP1 region and sequence comparison was performed with reference strains registered in Genbank. Male-to-female ratio confirmed approximately 5:4. The major clinical manifestation of patients infected with CVB1 was aseptic meningitis (55.8%). The other clinical symptoms were herpangina or hand-foot-mouth disease (22.1%) and neonatal sepsis (7.7%). The sequences of CVB1 isolates were divided into four genetic clusters (A-D) with at least 15% diversity between the clusters. Almost all the CVB1 isolates in Korea from 2008 to 2009 were in cluster D (except for 2 cases). The homology relationship was also similar between the Korean CVB1 strains and US strain (above 93%). It is possible that Korean CVB1 isolates found during 2008-2009 originated from the US strains found during 2006-2008. The identification of CVB1 in South Korea shows the potential of EVs to cause serious disease in an unpredictable fashion.


Assuntos
Infecções por Coxsackievirus/epidemiologia , Infecções por Coxsackievirus/virologia , Enterovirus/classificação , Enterovirus/isolamento & purificação , Adolescente , Criança , Pré-Escolar , Análise por Conglomerados , Enterovirus/genética , Feminino , Genótipo , Humanos , Lactente , Masculino , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , República da Coreia/epidemiologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Proteínas Estruturais Virais/genética
12.
J Korean Med Sci ; 28(1): 120-7, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23341722

RESUMO

In 2009, the first outbreak of hand, foot and mouth disease (HFMD) or herpangina (HP) caused by enterovirus 71 occurred in the Republic of Korea. This study inquired into risk factors associated with complications of HFMD or HP. A retrospective medical records review was conducted on HFMD or HP patients for whom etiologic viruses had been verified in 2009. One hundred sixty-eight patients were examined for this investigation. Eighty patients were without complications while 88 were accompanied by complications, and 2 had expired. Enterovirus 71 subgenotype C4a was the most prevalent in number with 67 cases (54.9%). In the univariate analysis, the disease patterns of HFMD rather than HP, fever longer than 4 days, peak body temperature over 39℃, vomiting, headache, neurologic signs, serum glucose over 100 mg/dL, and having an enterovirus 71 as a causative virus were significant risk factors of the complications. After multiple logistic analysis, headache (Odds ratio [OR], 10.75; P < 0.001) and neurologic signs (OR, 42.76; P < 0.001) were found to be the most significant factors. Early detection and proper management of patients with aforementioned risk factors would be necessary in order to attain a better clinical outcome.


Assuntos
Doença de Mão, Pé e Boca/complicações , Herpangina/complicações , Adolescente , Adulto , Glicemia/análise , Temperatura Corporal , Enterovirus Humano A/genética , Enterovirus Humano A/isolamento & purificação , Feminino , Febre/etiologia , Genótipo , Doença de Mão, Pé e Boca/virologia , Cefaleia/etiologia , Herpangina/virologia , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , República da Coreia , Estudos Retrospectivos , Fatores de Risco , Vômito/etiologia , Adulto Jovem
13.
Foodborne Pathog Dis ; 10(5): 461-6, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23627928

RESUMO

An epidemiological survey on human norovirus (NoV)-associated gastroenteritis was conducted to clarify the prevalence of NoV infections in children and adults in Korea. Recombinant capsid proteins from three major NoV genotypes (GI-4, GII-3, and GII-4) were expressed using a baculovirus expression system, and the morphology and antigenicity of self-assembled virus-like particles were then confirmed by electron microscopy and Western blotting with a NoV-specific antibody. To determine seroprevalence, an enzyme-linked immunosorbent assay was performed to detect antibodies against virus-like particles antigen in 346 serum specimens collected from persons who visited five public heath care centers for regular physical examination in Jeollanam-do, Korea, between 2005 and 2006. The seroprevalence of immunoglobulin G antibodies against the GI-4, GII-3, and GII-4 NoV genotypes was 84.1%, 76.3%, and 94.5%, respectively. A rapid decrease in seroprevalence occurred after birth, with the lowest levels observed in the <23-month age group, and a steep increase in seroprevalence occurred in early childhood, reaching 60.5% for GI-4, 65.1% for GII-3, and 90.7% for GII-4 at age 2-5 years, and over 80% for all three genotypes in subjects aged 20 years or older. The seroprevalence of different NoV genotypes statistically differed across the age groups (p<0.01).


Assuntos
Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Infecções por Caliciviridae/epidemiologia , Proteínas do Capsídeo/imunologia , Gastroenterite/epidemiologia , Norovirus/isolamento & purificação , Adolescente , Adulto , Antígenos Virais/genética , Baculoviridae/genética , Baculoviridae/metabolismo , Infecções por Caliciviridae/virologia , Proteínas do Capsídeo/genética , Criança , Pré-Escolar , Fezes/virologia , Feminino , Gastroenterite/virologia , Genótipo , Humanos , Imunoglobulina G/sangue , Lactente , Recém-Nascido , Pessoa de Meia-Idade , Norovirus/classificação , Norovirus/genética , Norovirus/imunologia , RNA Viral/genética , Proteínas Recombinantes , República da Coreia/epidemiologia , Estudos Soroepidemiológicos , Adulto Jovem
14.
Animals (Basel) ; 13(4)2023 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-36830388

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections have been frequently reported in companion dogs and cats worldwide during the ongoing coronavirus disease. However, RT-qPCR methods developed for humans have been used for the diagnosis of SARS-CoV-2 infections in suspected companion dogs and cats owing to the lack of the companion animal-tailored methods. Therefore, we developed a multiplex RT-qPCR (mRT-qPCR) using newly designed primers and probes targeting RdRp and N genes of all currently circulating SARS-CoV-2 variants as well as the canine or feline 16S rRNA gene as an endogenous internal positive control (EIPC) for reliable diagnosis of SARS-CoV-2 infection from suspected dogs and cats. The developed mRT-qPCR assay specifically detected the target genes of SARS-CoV-2 but no other canine or feline pathogens. Furthermore, canine and feline EIPCs were stably amplified by mRT-qPCR in samples containing canine- or feline-origin cellular materials. This assay has high repeatability and reproducibility, with an optimal limit of detection (<10 RNA copies per reaction) and coefficients of variation (<1.0%). The detection rate of SARS-CoV-2 of the developed mRT-qPCR was 6.6% for canine and feline nasopharyngeal samples, which was consistent with that of a commercial mRT-qPCR kit for humans. Collectively, the newly developed mRT-qPCR with canine and feline EIPC can efficiently diagnose and evaluate the viral load in field specimens and will be a valuable tool for etiological diagnosis, epidemiological study, and controlling SARS-CoV-2 infections in canine and feline populations.

15.
J Clin Microbiol ; 50(4): 1252-7, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22238447

RESUMO

Hepatitis A virus is one of the most prominent causes of fecally transmitted acute hepatitis worldwide. In order to characterize the viral agents causing an outbreak in Korea (comprising North and South Korea) from June 2007 to May 2009, we collected specimens and performed genotyping of the VP1/P2A and VP3/VP1 regions of hepatitis A virus. We then used a multiple-alignment algorithm to compare the nucleotide sequences of the 2 regions with those of reference strains. Hepatitis A virus antibodies were detected in 64 patients from 5 reported outbreaks (North Korea, June 2007 [n = 11]; Jeonnam, April 2008 [n = 15]; Daegu, May 2008 [n = 13]; Seoul, May 2009 [n = 22]; and Incheon, May 2009 [n = 3]). We found 100% homology between strains isolated from the Kaesong Industrial Region and Jeonnam. While those strains were classified as genotype IA strains, strains from Seoul and Incheon were identified as genotype IIIA strains and showed 98.9 to 100% homology. Genotype IIIA was also dominant in Daegu, where strains were 95.7 to 100% homologous. All hepatitis A virus strains isolated from the Kaesong Industrial Region, Jeonnam, Seoul, and Incheon belonged to a single cluster. However, strains from Daegu could be classified into 2 clusters, suggesting that the outbreak had multiple sources. This study indicates that hepatitis A virus strains of 2 different genotypes are currently cocirculating in Korea. Moreover, it documents an increasing prevalence of genotype IIIA strains in the country.


Assuntos
Epidemias , Vírus da Hepatite A Humana/genética , Hepatite A/virologia , Proteínas Estruturais Virais/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Feminino , Genótipo , Hepatite A/epidemiologia , Humanos , Masculino , Dados de Sequência Molecular , Tipagem Molecular , Filogenia , República da Coreia/epidemiologia , Análise de Sequência de RNA
16.
Virol J ; 9: 38, 2012 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-22336050

RESUMO

BACKGROUND: Enteroviruses (EVs) are the leading cause of aseptic meningitis, which is the most frequent central nervous system infection worldwide. We aimed to characterize the EVs involved in an aseptic meningitis outbreak in Korea in 2008. In Korea, Echovirus type 30 (E30) and E6 have been associated with outbreaks and frequent meningitis. METHODS: During 2008, through nationwide surveillance, we collected specimens from 758 patients with aseptic meningitis-related clinical manifestations. The detection of EVs from specimens was subjected to a diagnostic real-time RT-PCR in the 5' NCR. A semi-nested polymerase chain reaction (PCR) to amplify sequences from the VP1 region and sequence comparison with reference strains registered in Genbank was performed for the genotype determination. RESULTS: Most patients (98%) in this outbreak were children < 15 years of age. The temporal distribution of the E6 and E30 epidemics showed an obvious seasonal pattern during the short period from June to July. A large majority of the EV-positive patients experienced fever, headache, vomiting, and neck stiffness. Some patients also showed cold symptoms, sore throat, altered mental status, and seizures. We did not observe a higher fatality rate in children with E6 or E30 infection. Most of the patients recovered uneventfully. In most cases, the cerebrospinal fluid (CSF) profile was studied, and generally showed a higher than normal white blood cell count (≥ 5/mm(3)). We detected EVs from 513 patients (67.68%) and identified the EV genotype in 287 patients. E30 (n = 155, 50.4%) and E6 (n = 95, 33.1%) were the predominant genotypes. E9, E1, E7, E16, coxsackievirus A3, 4, 6, coxsackievirus B1, 3, and 10 were also identified. According to phylogenetic analysis, E30 belonged to subgroup 4b, and E6, to the C4 subgroup. CONCLUSIONS: Conclusively, aseptic meningitis was the most common manifestation in children with either echovirus 30 or 6 infection. Identification of E6 and E30 as the prominent EVs in the 2008 outbreak in South Korea shows the potential of EVs to cause a serious disease in an unpredictable (fashion. Our findings provide new) insights into the clinical and virological features of the aseptic meningitis outbreak caused by E30 and E6.


Assuntos
Infecções por Echovirus/diagnóstico , Infecções por Echovirus/epidemiologia , Enterovirus Humano B/isolamento & purificação , Meningite Viral/diagnóstico , Meningite Viral/epidemiologia , Adolescente , Criança , Pré-Escolar , Infecções por Echovirus/virologia , Enterovirus Humano B/classificação , Enterovirus Humano B/genética , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Meningite Viral/virologia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , República da Coreia/epidemiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Virologia/métodos
17.
Virol J ; 8: 330, 2011 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-21714898

RESUMO

BACKGROUND: Enterovirus (EV) infections are commonly associated with encephalitis and meningitis. Detection of enteroviral RNA in clinical specimens has been demonstrated to improve the management of patients, by ruling out other causes of disease. METHOD: To develop a sensitive and reliable assay for routine laboratory diagnosis, we developed a real-time one step reverse transcription polymerase chain reaction (RT-PCR) assay with minor groove binder probes and primers modified with complementary locked primer technology (TMC-PCR). We checked the sensitivity of the developed assay by comparing it to a previously published TaqMan probe real-time one-step RT-PCR (TTN-PCR) procedure using enteroviral isolates, Enterovirus Proficiency panels from Quality Control on Molecular Diagnostics (QCMD-2007), and clinical specimens from patients with suspected EV infections. RESULTS: One hundred clinical specimens from 158 suspected viral meningitis cases were determined to be positive by the TMC-PCR assay (63.29%), whereas only 60 were found to be positive by the TTN-PCR assay (37.97%). The positive and negative agreements between the TMC-PCR and TTN-PCR assays were 100% and 59.2%, respectively. CONCLUSION: This data suggest that the TMC-PCR assay may be suitable for routine diagnostic screening from patient suspected EV infection.


Assuntos
Primers do DNA/genética , Infecções por Enterovirus/diagnóstico , Enterovirus/genética , Enterovirus/isolamento & purificação , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Virologia/métodos , Enterovirus/classificação , Infecções por Enterovirus/virologia , Humanos , Sensibilidade e Especificidade
18.
Virol J ; 8: 79, 2011 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-21345236

RESUMO

An outbreak of echovirus 5 (ECV 5) occurred in Korea in 2006, marking the first time this virus had been identified in the country since enterovirus surveillance began in 1993. Using a sample isolated from a young male patient with aseptic meningitis, we performed sequencing of the Korean ECV 5 strain and compared it with a prototype strain (Noyce). At the nucleotide level, the P1 region (85.3%) had the highest identity value; at the amino acid level, the P3 region (98.0%) had the highest identity value. The two strains shared all cleavage sites, with the exception of the VP1/2A site, which was TY/GA in the Noyce strain but TR/GA in the Korean ECV 5 isolate. In Vero cells infected with the Korean ECV 5 isolate, no cytotoxicity was observed in the presence of azidothymidine, acyclovir, amantadine, lamivudine, or ribavirin, when the drugs were administered at a CC50 value >100 µg/mL. Of the five drugs, only amantadine (IC50: 1 ± 0.42 µg/mL, TI: 100) and ribavirin (IC50: 22 ± 1.36 µg/mL, TI: 4.55) had any antiviral activity against the Korean ECV 5 isolate.


Assuntos
Antivirais/farmacologia , Infecções por Echovirus/virologia , Enterovirus Humano B/efeitos dos fármacos , Enterovirus Humano B/genética , Variação Genética , Infecções por Echovirus/tratamento farmacológico , Infecções por Echovirus/epidemiologia , Enterovirus Humano B/classificação , Enterovirus Humano B/isolamento & purificação , Humanos , Coreia (Geográfico)/epidemiologia , Filogenia , Proteínas Virais/genética
19.
J Pediatr Gastroenterol Nutr ; 52(3): 314-8, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21150655

RESUMO

OBJECTIVE: Pneumatosis intestinalis (PI) in children is associated with immunosuppression, mucosal disruption from trauma, obstructive pulmonary disease, congenital heart disease, and gastrointestinal infections. Our study is the first report of norovirus infection-associated PI. PATIENTS AND METHODS: A retrospective review was performed in pediatric patients (older than 30 days) with PI from March 2005 to April 2009. Since December 2008, in addition to routine stool examinations, reverse-transcriptase polymerase chain reaction testing for calicivirus (norovirus and sapovirus), adenovirus, astrovirus, and enterovirus has been performed. RESULTS: Twenty-seven patients with PI were identified. The median age was 1.4 (range 0.2-14.8 years). Seventeen patients (63.0%) were immunocompromised hosts. Pathogens were identified in 5 immunocompromised patients (5/27 and 5/8 since December 2008). Of note, norovirus was identified in 4 patients (80%, 4/5) during the cold weather season. The genotype of noroviruses in these patients was GII-4. Among 27 patients with PI, 10 patients (37.0%) developed PI in the spring and 11 (40.7%) in the winter. Twenty-four patients survived (88.9%, 24/27). None of the patients with norovirus or rotavirus infection died. CONCLUSIONS: Our data suggest that norovirus infection may contribute to the development of PI in immunocompromised hosts.


Assuntos
Infecções por Caliciviridae/complicações , Hospedeiro Imunocomprometido , Norovirus , Pneumatose Cistoide Intestinal/virologia , Adolescente , Infecções por Caliciviridae/virologia , Criança , Pré-Escolar , Temperatura Baixa , Feminino , Genótipo , Humanos , Terapia de Imunossupressão/efeitos adversos , Lactente , Masculino , Norovirus/genética , Estudos Retrospectivos , Estações do Ano
20.
Emerg Infect Dis ; 16(11): 1764-6, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21029539

RESUMO

We assessed neurologic sequelae associated with an enterovirus 71 (EV71) outbreak in South Korea during 2009. Four of 94 patients had high signal intensities at brainstem or cerebellum on magnetic resonance imaging. Two patients died of cardiopulmonary collapse; 2 had severe neurologic sequelae. Severity and case-fatality rates may differ by EV71 genotype or subgenotype.


Assuntos
Doenças do Sistema Nervoso Central/epidemiologia , Doenças do Sistema Nervoso Central/etiologia , Enterovirus Humano A , Infecções por Enterovirus/complicações , Infecções por Enterovirus/epidemiologia , Criança , Pré-Escolar , Surtos de Doenças , Enterovirus Humano A/genética , Infecções por Enterovirus/mortalidade , Feminino , Genótipo , Humanos , Lactente , Masculino , Dados de Sequência Molecular , República da Coreia/epidemiologia
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