RESUMO
We compared rapid antigen detection kits widely used for the rapid diagnosis of group A streptococcal pharyngitis, evaluating their minimum detection sensitivity and operability in five levels. Five kits based on the immunochromatographic method were used: ImunoAce Strep A (Tauns), ImunoAce Strep A Neo (Tauns), Quick Navi-StrepA2 (Denka), Quick Vue Dipstick Strep A (SB Bioscience) and RapidTesta Strep A (SEKISUI MEDICAL). Thirteen strains were tested: 10 clinical isolates of Streptococcus pyogenes, 2 strains of Streptococcus dysgalactiae subsp. equisimilis (SDSE), and S. pyogenes ATCC 19615. All kits had the same or higher minimum detection sensitivity than previously reported. ImunoAce StrepA Neo had the highest detection sensitivity and the best overall evaluation among the group A streptococcal rapid antigen detection kits used in this study. The detection sensitivity of SDSE with group A polysaccharide antigen was comparable to that of S. pyogenes. Although culture tests are necessary to confirm the causative organism, SDSE may present with clinical symptoms similar to those of S. pyogenes, and detection with a rapid antigen detection kit may be of therapeutic value.
Assuntos
Faringite , Infecções Estreptocócicas , Humanos , Streptococcus pyogenes , Infecções Estreptocócicas/diagnóstico , Faringite/diagnóstico , Faringite/microbiologia , Kit de Reagentes para DiagnósticoRESUMO
We investigated the efficiency of the Verigene Enteric Pathogens Nucleic Acid Test (Verigene EP test), which is an automated microarray-based assay system that enables rapid and simultaneous genetic detection of gastrointestinal pathogens and toxins, including those in the Campylobacter Group, Salmonella species, Shigella species, the Vibrio Group, Yersinia enterocolitica, Shiga toxin 1 and 2, norovirus GI/GII, and rotavirus A. Three clinical laboratories evaluated the Verigene EP test, using 268 stool samples for bacterial and toxin genes and 167 samples for viral genes. Culture-based reference methods were used for the detection of bacteria and toxins, while a different molecular assay was used for viral detection. The overall concordance rate between the Verigene EP test and the reference methods for the 1940 assays was 99.0%. The overall sensitivity and specificity of the Verigene EP test were 97.0% and 99.3%, respectively. Of the 19 samples with discordant results, 13 samples were false positives and six were false negatives. The Verigene EP test simultaneously detected two targets in 11 samples; overall, the test demonstrated high efficiency in detecting crucial diarrheagenic pathogens, indicating its suitability for clinical practice.
Assuntos
Toxinas Bacterianas/isolamento & purificação , Diarreia/diagnóstico , Gastroenterite/microbiologia , Microbioma Gastrointestinal , Toxinas Bacterianas/genética , Diarreia/genética , Diarreia/microbiologia , Fezes/microbiologia , Gastroenterite/diagnóstico , Gastroenterite/genética , Humanos , Técnicas de Diagnóstico Molecular , Norovirus/genética , Norovirus/isolamento & purificação , Norovirus/patogenicidade , Técnicas de Amplificação de Ácido Nucleico/métodos , Toxina Shiga I/química , Toxina Shiga I/genética , Toxina Shiga I/isolamento & purificação , Shigella/genética , Shigella/isolamento & purificação , Shigella/patogenicidadeRESUMO
Recently, a new rapid assay for the detection of tcdB gene of Clostridioides difficile was developed using the GENECUBE. The assay can directly detect the tcdB gene from stool samples without a purification in approximately 35 minutes with a few minutes of preparation process. We performed a prospective comparative study of the performance of the assay at eight institutions in Japan. Fresh residual stool samples (Bristol stool scale ≥5) were used and comparisons were performed with the BD MAX Cdiff assay and toxigenic cultures. For the evaluation of 383 stool samples compared with the BD MAX Cdiff assay, the sensitivity, and specificity of the two assays was 99.0% (379/383), 98.1% (52/53), 99.1% (327/330), respectively. In the comparison with toxigenic culture, the total, sensitivity, and specificity were 96.6% (370/383), 85.0% (51/60), and 98.8% (319/323), respectively. The current investigation indicated the GENECUBE Clostridioides difficile assay has equivalent performance with the BD MAX Cdiff assay for the detection of tcdB gene of C. difficile.
Assuntos
Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Clostridioides difficile/genética , Fezes/microbiologia , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/diagnóstico , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Enterotoxinas/genética , Humanos , Reação em Cadeia da Polimerase/métodos , Kit de Reagentes para Diagnóstico , Proteínas Repressoras/genética , Sensibilidade e EspecificidadeRESUMO
The prevalence of bovine foamy virus (BFV) infections in cattle on farms in the Kanto region of Japan was determined using agar gel immunodiffusion (AGID) test and polymerase chain reaction (PCR). Six out of 20 farms contained BFV-positive cattle. Furthermore, 16.7% (91/545) of all cattle tested positive for BFV. This suggested that BFV-infected cattle are widely prevalent in Japan. Positive results for BFV infection were consistent between AGID and PCR tests. Additionally, we tested for bovine leukemia virus (BLV) infections at nine farms, primarily those containing BFV-infected cows. At each farm, the infection rate of BFV was lower than that of BLV. Further, cattle that were PCR-positive but antibody-negative, indicating immune tolerance to BFV, were not detected.
Assuntos
Doenças dos Bovinos/virologia , Leucose Enzoótica Bovina/epidemiologia , Infecções por Retroviridae/veterinária , Animais , Anticorpos Antivirais/análise , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/epidemiologia , DNA Viral/análise , Leucose Enzoótica Bovina/sangue , Imunodifusão/veterinária , Japão/epidemiologia , Vírus da Leucemia Bovina/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Infecções por Retroviridae/sangue , Infecções por Retroviridae/epidemiologia , Spumavirus/isolamento & purificaçãoRESUMO
For understanding the factors affecting bovine viral diarrhea virus (BVDV) transmission, this study investigated the distribution of BVDV and the epidemiological features of persistently infected (PI) cattle in Ibaraki Prefecture of Japan, and identified farm-level risk factors associated with BVDV infection, with a focus on within-farm transmission and PI animal detection. Among all 377 dairy farms, forty-four PI cattle were identified on 22 farms. Thirty-eight and six PI cattle were born on their current farms or purchased, respectively. Twenty-six PI cattle were born from pregnancies on their current farms, seven from pregnancies in summer pastures, and eight from pregnancies on other farms. The within-farm seroprevalence on farms with PI animals was significantly higher than that on farms without PI cattle. Of 333 farms holding homebred cattle without movement records, antibody-positivity in homebred cattle was observed on 194 farms; these cattle were likely infected by within-farm transmission. Herd size, summer pasturing, and BVDV infection status of the nearest dairy farm were risk factors associated with within-farm transmission. Likewise, herd size, summer pasturing, and the proportion of purchased cattle were related to PI animal occurrence. This study shows the risk of within-farm transmission and occurrence of PI animals after the introduction of BVDV via purchasing and summer pasturing, and illustrates the significant role of PI cattle in circulating BVDV. More effective measures for screening BVDV infection and PI animals, including intensive tests targeting moved cattle and newborn calves, and bulk milk surveillance, are required to control the spread of BVDV in Japan.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina Tipo 1 , Vírus da Diarreia Viral Bovina , Fazendas , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Doença das Mucosas por Vírus da Diarreia Viral Bovina/transmissão , Bovinos , Feminino , Incidência , Japão/epidemiologia , Gravidez , Fatores de Risco , Estações do Ano , Estudos SoroepidemiológicosRESUMO
An 83-year-old previously self-sufficient man was referred to our hospital for a fever, severe tenderness over the lumbar spine, and elevated C-reactive protein levels. Computed tomography revealed fluid collection in the intervertebral space of L3/4. Gram-positive, short rod-shaped bacteria were isolated from two sets of blood cultures. A 16S rRNA sequence analysis of an isolate showed a similarity of 98.1% to the nearest type strain Brachybacterium squillarum JCM 16464T. Biochemical characteristics of the presently isolated strain differed from those of the most closely related species of the genus Brachybacterium. The patient was successfully discharged on day 73 of admission with antimicrobial therapies and showed no recurrence during outpatient visits. Brachybacterium spp. have mainly been isolated from the environment, and human Brachybacterium infections have rarely been documented to date. To our knowledge, this is the first clinical isolation of Brachybacterium sp. as a causative pathogen of bloodstream infection.
Assuntos
Infecções por Actinomycetales/microbiologia , Bacteriemia/microbiologia , Vértebras Lombares/patologia , Micrococcaceae/isolamento & purificação , Infecções por Actinomycetales/sangue , Infecções por Actinomycetales/diagnóstico , Infecções por Actinomycetales/tratamento farmacológico , Idoso de 80 Anos ou mais , Anti-Infecciosos/uso terapêutico , Bacteriemia/sangue , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Biópsia por Agulha , Proteína C-Reativa/análise , Creatinina/análise , DNA Bacteriano/genética , Humanos , Masculino , Micrococcaceae/genética , RNA Ribossômico 16S/genéticaRESUMO
From 29 November 2016 to 24 January 2017, sixty-three cases of H5N6 highly pathogenic avian influenza virus (HPAIV) infections were detected in wild birds in Ibaraki Prefecture, Japan. Here, we analyzed the genetic, temporal, and geographic correlations of these 63 HPAIVs to elucidate their dissemination throughout the prefecture. Full-genome sequence analysis of the Ibaraki isolates showed that 7 segments (PB2, PB1, PA, HA, NP, NA, NS) were derived from G1.1.9 strains while the M segment was from G1.1 strains; both groups of strains circulated in south China. Pathological studies revealed severe systemic infection in dead swans (the majority of dead birds and the only species necropsied), thus indicating high susceptibility to H5N6 HPAIVs. Coalescent phylogenetic analysis using the 7 G1.1.9-derived segments enabled detailed analysis of the short-term evolution of these highly homologous HPAIVs. This analysis revealed that the H5N6 HPAIVs isolated from wild birds in Ibaraki Prefecture were divided into 7 groups. Spatial analysis demonstrated that most of the cases concentrated around Senba Lake originated from a single source, and progeny viruses were transmitted to other locations after the infection expanded in mute swans. In contrast, within just a 5-km radius of the area in which cases were concentrated, three different intrusions of H5N6 HPAIVs were evident. Multi-segment analysis of short-term evolution showed that not only was the invading virus spread throughout Ibaraki Prefecture but also that, despite the small size of this region, multiple invasions had occurred during winter 2016-2017.
Assuntos
Aves/virologia , Genoma Viral , Vírus da Influenza A/patogenicidade , Influenza Aviária/transmissão , Influenza Aviária/virologia , Animais , Animais Selvagens/virologia , Galinhas/virologia , Patos/virologia , Vírus da Influenza A/genética , Influenza Aviária/epidemiologia , Influenza Aviária/mortalidade , Japão/epidemiologia , Filogenia , Vírus Reordenados/genética , Vírus Reordenados/patogenicidade , Análise de Sequência de DNARESUMO
A 56-year-old previously healthy man was hospitalized due to a 10-day history of neck pain and an elevated C-reactive protein level. Gram-negative spiral bacilli were isolated from his blood, and Helicobacter cinaedi was confirmed using 16S rRNA sequencing. The infectious focus was not identified by initial cervical magnetic resonance imaging (MRI); however, repeated MRI demonstrated prominent high signal intensity in the entire region of the C6-C7 vertebrae and C6/C7 disc space. Furthermore, fluorodeoxyglucose-positron emission tomography/computed tomography showed no significant uptake, other than in the C6-C7 region. The patient was successfully treated with ceftriaxone for six weeks without sequelae.
Assuntos
Vértebras Cervicais/microbiologia , Infecções por Helicobacter/tratamento farmacológico , Hospedeiro Imunocomprometido , Osteomielite/microbiologia , Proteína C-Reativa/análise , Ceftriaxona/uso terapêutico , Diagnóstico Diferencial , Helicobacter/genética , Infecções por Helicobacter/genética , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Osteomielite/tratamento farmacológico , RNA Ribossômico 16S/genética , Tomografia Computadorizada por Raios XRESUMO
The Verigene Gram-positive blood culture test (BC-GP) and the Verigene Gram-negative blood culture test (BC-GN) identify representative Gram-positive bacteria, Gram-negative bacteria and their antimicrobial resistance by detecting resistance genes within 3 h. Significant benefits are anticipated due to their rapidity and accuracy, however, their clinical utility is unproven in clinical studies. We performed a clinical trial between July 2014 and December 2014 for hospitalized bacteremia patients. During the intervention period (N = 88), Verigene BC-GP and BC-GN was used along with conventional microbiological diagnostic methods, while comparing the clinical data and outcomes with those during the control period (N = 147) (UMIN registration ID: UMIN000014399). The median duration between the initiation of blood culture incubation and the reporting time of the Verigene system results was 21.7 h (IQR 18.2-26.8) and the results were found in 88% of the cases by the next day after blood cultures were obtained without discordance. The hospital-onset infection rate was higher in the control period (24% vs. 44%, p = 0.002), however, no differences were seen in co-morbidities and severity between the control and intervention periods. During the intervention period, the time of appropriate antimicrobial agents' initiation was significantly earlier than that in the control period (p = 0.001) and most cases (90%; 79/88) were treated with antimicrobial agents with in-vitro susceptibility for causative bacteria the day after the blood culture was obtained. The costs for antimicrobial agents were lower in the intervention period (3618 yen vs. 8505 yen, p = 0.001). The 30-day mortality was lower in the intervention period (3% vs. 13%, p = 0.019).
Assuntos
Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Técnicas de Diagnóstico Molecular/instrumentação , Idoso , Antibacterianos/uso terapêutico , Bacteriemia/tratamento farmacológico , Doenças Transmissíveis/diagnóstico , Doenças Transmissíveis/tratamento farmacológico , Doenças Transmissíveis/microbiologia , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , Feminino , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/genética , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/genética , Humanos , Masculino , Análise em Microsséries/métodos , Estudos ProspectivosRESUMO
The Verigene Gram-Negative Blood Culture Nucleic Acid Test (BC-GN) is a microarray-based assay that enables rapid detection of 9 common Gram-negative bacteria and 6 resistance determinants directly from positive blood cultures. We compared the performance of BC-GN with currently used automated systems, testing 141 clinical blood cultures and 205 spiked blood cultures. For identification of BC-GN target organisms in clinical and spiked blood cultures, the BC-GN assay showed 98.5% (130/132) and 98.9% (182/184) concordance, respectively. Of 140 resistance genes positively detected in clinical and spiked blood cultures with the BC-GN test, 139 (99.3%) were confirmed by PCR, and the detection results were consistent with the resistance phenotypes observed. The BC-GN assay, thus, can potentially improve care for sepsis patients by enabling timely detection and targeted antimicrobial therapy.
Assuntos
Bacteriemia/diagnóstico , Bacteriemia/microbiologia , DNA Bacteriano/análise , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/isolamento & purificação , Testes de Sensibilidade Microbiana/métodos , Técnicas de Diagnóstico Molecular/métodos , Automação Laboratorial/métodos , DNA Bacteriano/genética , Bactérias Gram-Negativas/classificação , Humanos , Fatores de TempoRESUMO
A 43-year-old man was referred to our hospital for an acute-onset fever and left flank pain. He had been previously diagnosed with lymphangioma, and abdominal computed tomography showed pararenal cysts with fat stranding around the left kidney, of which infection was subsequently confirmed on magnetic resonance imaging. Gram-negative spiral bacilli were isolated from two sets of blood cultures, and Helicobacter cinaedi was identified using 16S rRNA sequencing. The patient was successfully treated with ceftriaxone therapy without recurrence. A multilocus sequence typing analysis indicated the current H. cinaedi strain differed from previous strains isolated in Japan.
Assuntos
Bacteriemia/diagnóstico , Infecções por Helicobacter/diagnóstico , Linfocele/diagnóstico , Adulto , Bacteriemia/tratamento farmacológico , Bacteriemia/microbiologia , Ceftriaxona/uso terapêutico , Helicobacter/genética , Helicobacter/isolamento & purificação , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Humanos , Japão , Linfocele/diagnóstico por imagem , Masculino , Tipagem de Sequências Multilocus , RNA Ribossômico 16S/genética , RadiografiaRESUMO
Human pinworms, Enterobius vermicularis, are normally recognized as minor pathogens. However, a fatal case of human pinworm infection has been reported in a nonhuman primate, a zoo reared chimpanzee. Here, we histopathologically examined the lesions in tissues from the deceased chimpanzee and genetically characterized the isolated worms to investigate the pathogenicity and determine the phylogeny. We identified ulcers deep in the submucosa where many parasites were found to have invaded the lamina propria mucosa or submucous tissue. An inflammatory reaction consisting mainly of neutrophils and lymphocytes but not eosinophils was observed around the parasites, and intense hemorrhage in the lamina propria was confirmed. The parasites were morphologically similar to E. vermicularis based on the shape of the copulatory spicules. Mitochondrial cytochrome c oxidase subunit 1 gene products were amplified from worm DNA by PCR and were genetically identified as E. vermicularis based on >98.7% similarity of partial sequences. Phylogenetic analysis revealed that the sequences clustered together with other chimpanzee E. vermicularis isolates in a group which has been referred to as type C and which differs from human isolates (type A). The samples were negative for bacterial pathogens and Entamoeba histolytica indicating that E. vermicularis could be pathogenic in chimpanzees. Phylogenetic clustering of the isolates indicated that the parasite may be host specific.
Assuntos
Colite/parasitologia , Enterobíase/veterinária , Enterobius/genética , Pan troglodytes/parasitologia , Animais , Colite/patologia , Colo/parasitologia , Colo/patologia , DNA de Helmintos/genética , Enterobíase/parasitologia , Enterobius/isolamento & purificação , Feminino , Filogenia , Reação em Cadeia da PolimeraseRESUMO
A ten-year-old Shetland pony gelding showed low appetite, ataxia, peculiar swaying, clouding of consciousness, and ultimately died. At necropsy, multiple coalescing granulomatous foci were detected in the kidneys, and small necrotic lesions were found in the cerebellum. Histologic examination of the renal tissue sections revealed extensive granuloma, and Halicephalobus gingivalis-like nematodes were seen. Similar nematodes were found in the granulomatous or necrotic lesions of the renal lymph nodes and cerebellum, and were also frequently detected in cerebrospinal meningovascular lesions. Morphologic features together with partial ribosomal RNA gene sequences of the nematodes in the lesions revealed that they were H. gingivalis. The present results indicated that H. gingivalis caused granulomatous nephritis and meningoencephalomyelitis in this pony gelding.