Rinovirus y metapneumovirus en pacientes con infección respiratoria aguda grave / Rhinovirus and metapneumovirus in patients with severe acute respiratory infection

Introducción. Los virus son los principales agentes etiológicos en las infecciones respiratorias agudas graves; un alto porcentaje queda sin diagnóstico viral. Objetivo. Describir la frecuencia de rinovirus y metapneumovirus en pacientes pediátricos de una unidad centinela de Mar del Plata con infección respiratoria aguda grave y resultado negativo para virus clásicos por inmunofluorescencia y biología molecular. Población y métodos. Se realizó un estudio descriptivo de corte transversal. Se evaluó la presencia de rinovirus y metapneumovirus por biología molecular en 163 casos negativos para panel respiratorio por técnicas de vigilancia referencial, durante todo el año 2015. Resultados. Se detectó rinovirus en el 51,5 % de los casos, metapneumovirus en el 9,8 % y coinfección rinovirus-metapneumovirus en el 6,1 %. Fueron negativos para ambos virus el 32,5 %. Conclusiones. La selección de muestras sin diagnóstico virológico permitió identificar rinovirus y metapneumovirus como agentes causales de infecciones respiratorias agudas graves pediátricas y su impacto en la morbimortalidad infantil y en nuestro sistema sanitario.

Introduction. Viruses are the main etiologic agents involved in severe acute respiratory tract infections; a viral diagnosis is not established in a high percentage of cases. Objective. To describe the frequency of rhinovirus and metapneumovirus in pediatric patients with severe acute respiratory infection and negative results for typical viruses by immunofluorescence and molecular biology at a sentinel unit of Mar del Plata. Population and methods. This was a descriptive, cross-sectional study. The presence of rhinovirus and metapneumovirus was assessed by molecular biology in 163 cases negative for respiratory panel by referral surveillance techniques throughout 2015. Results. Rhinovirus was detected in 51.5% of cases, metapneumovirus in 9.8%, and coinfection with rhinovirus and metapneumovirus in 6.1%. Results were negative for both viruses in 32.5%. Conclusions. The selection of samples without a viral diagnosis allowed us to identify rhinovirus and metapneumovirus as causative agents of severe acute respiratory infections in children and assess their impact on child morbidity and mortality and on our health care system

Asociación temporal entre la circulación de virus respiratorios y hospitalizaciones por enfermedad de Kawasaki / Temporal association between the circulation of respiratory viruses and hospitalizations due to Kawasaki disease

INTRODUCCIÓN: La etiología de la enfermedad de Kawasaki (EK) es desconocida, planteándose que infecciones virales la gatillan en pacientes susceptibles. OBJETIVO: Estudiar la asociación temporal entre la circulación de virus respiratorios y hospitalizaciones por EK en la Región Metropolitana (RM), Chile, entre 2010-2017. METODOLOGÍA: Estudio ecológico retrospectivo de casos de EK en pacientes bajo 18 años de edad, en base a egresos hospitalarios. La circulación de virus se analizó mediante el reporte de la red de vigilancia metropolitana. Se utilizaron promedios móviles para EK (PMEK) y virus respiratorios (PMVR). RESULTADOS: Se registraron 14.902 casos de infecciones virales respiratorias entre 2010-2017. Se observó correlación directa entre PMVR-virus respiratorio sincicial (VRS) de un mes y año y PMEK del mes subsiguiente (coeficiente de correlación (ρ) = +0,441; p < 0,001), y una asociación similar para PMVR-influenza A (FLU A) (ρ = +0,362; p < 0,001). PMVR-influenza B (FLU B) y PMVR-metapneumovirus (MPV) presentan correlaciones directas con PMEK (ρ = +0,443; p < 0,001 y ρ = +0,412; p < 0,001, respectivamente), siendo contemporáneo en mes y año con EK para FLU B, mientras que MPV presenta un desfase de un mes entre PMVR y PMEK. CONCLUSIÓN: Existe correlación temporal directa entre la circulación de VRS, FLU A, FLU B y MPV con EK en niños de la RM, Chile.

BACKGROUND: The etiology of Kawasaki disease (KD) is unknown. It is believed that viral infections could trigger the disease in susceptible patients. AIM: To study the temporal association between the circulation of respiratory viruses and KD hospitalizations in the Metropolitan Region (MR), Chile, between 2010-2017. METHODS: Ecologic study consisting of a review of KD cases in children under 18 years of age based on hospital discharges. The circulation of respiratory viruses was analyzed using the report of the metropolitan surveillance network. Moving averages for KD (MAKD) and respiratory viruses (MARV) were used. RESULTS: 14,902 cases of respiratory virus infections were recorded between 2010-2017. A direct correlation was found between MARV-respiratory syncytial virus (RSV) of one month and year and MAKD of the subsequent month (correlation coefficient (ρ) = +0.441; p < 0.001). A similar association was found for MARV-influenza A (FLU A) (ρ = + 0.362; p < 0.001). MARV-influenza B (FLU B) and MARV-metapneumovirus (MPV) had direct correlations with MAKD (ρ = +0.443; p < 0.001 and ρ = +0.412; p < 0.001, respectively), being FLU B contemporary in month and year with KD, and MPV presenting a one-month lag. CONCLUSION: There is a direct temporal correlation between RSV, FLU A, FLU B and MPV circulation and KD in children from RM, Chile.

In vitro and in vivo anti-HMPV activity of JH001 isolated from Chinese medicinal herbs / 中国药理学与毒理学杂志

OBJECTIVE Human metapneumovirus (hMPV) is semblable to respiratory syncytial virus (RSV) which causes respiratory infections typically characterized by cough, runny nose, fever, and nasal congestion but sometimes progressing to bronchiolitis and pneumonia. Whereas, there is no corresponding drug to inhabit the virus. Studies of new compounds with potential anti-HMPV activity could produce clinical value. Chinese herbal medicine played a great role during COVID-19, therefore we choose some small molecular (JH001) extracted from botany to investigate therapeutic effect on hMPV and the underlying mechanisms. METHODS In this study, 16HBE cells were used as a model to explore in vitro antiviral effect. Cytotoxicity assays were performed before the antiviral tests, cell viability of 16HBE cells handled by different concentration of JH001 was estimated by Cell Counting Kit-8 (CCK-8). Then RT-qPCR, immunofluores?cence, and flow cytometer were used to test the viral titer after cells infected with hMPV. Eventually, 6-8 weeks mice were infected intranasally with 60 μL of hMPV, the control group was treated with 0.9％ saline water, other groups were administered with JH001 and ribavirin, then the lung virus titer and protective effect in lung were judged. RESULTS The obtained JH001 exhibited no cytotoxicity to 16HBE cells during 6.25 - 200 μmol · L-1. RT-QPCR demonstrated that JH001 showed obvious inhabitation to the viral replication and showed great significance compared with saline. And fluo?rescence exhibited distinct decrease of hMPV-N protein, flow cytometer results showed that MFI decrease evidently. Sig?nificant reduction of N-gene expression was observed in those mice treated with JH001 compared with saline group, which indicated that JH001 probably had protective and therapeutic effect on viral replication. CONCLUSION This study illustrated that JH001 might be a promising option for small molecular against hMPV and JH001 might be worthy of fur?ther development and used as a potential therapeutic strategy for other respiratory viruses in the future.

Human metapneumovirus-associated community-acquired pneumonia in adults during the first wave of COVID-19 / Journal of Rural Medicine

Objective: The clinical course of human metapneumovirus (hMPV) infection is similar to that of coronavirus 2019 disease (COVID-19). However, community-acquired hMPV infections in adults have not yet been sufficiently investigated. We examined the detection status of hMPV antigens and the clinical features of positive patients during the first wave of COVID-19, which coincided with the epidemic season of hMPV infection in Japan.Methods: In this cross-sectional, observational, and single-center study, we recruited consecutive individuals who visited the Japan Agricultural Cooperatives Kochi Hospital due to fever, respiratory symptoms, or close contact with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected persons during the period from January to May 2020.Results: The positive rate of immunochromatography for hMPV antigens from nasopharyngeal swabs was 9.5% (4/42), and four positive cases were community-acquired pneumonia (CAP) (5.3% of all CAP). The positive rate of hMPV antigens in the CAP group (30.8%, 4/13) was higher than that in the non-pneumonia group (0.0%, 0/19) (p < 0.05). The average age of the four adult patients with CAP was 69.8 years (range 35–93). Mean white blood cell counts and C-reactive protein blood levels were 6,250 cells/μL (3,500–12,180) and 4.30 mg/dL (4.05–7.04), respectively. Chest computed tomography images were diverse and two patients showed dense consolidation. No multi-organ disorder was noted during the clinical course in any of the four cases, and their prognoses were good.Conclusion: hMPV infection may be considered in the differential diagnosis of COVID-19 and CAP in Japan under the preventive measures for SARS-CoV-2 infection, at least during the epidemic season of hMPV infection.

Viral infection and epidemiological characteristics of human metapneumovirus in febrile respiratory syndrome cases in nine provinces in China from 2009 to 2021 / 中华实用儿科临床杂志

Objective:To clarify the infection and epidemic characteristics of the human metapneumovirus (HMPV) in Chinese patients with febrile respiratory syndrome (FRS), and to provide important baseline data for clinical diagnosis, treatment, prevention and control of HMPV-induced respiratory tract diseases in China.Methods:FRS cases from January 2009 to June 2021 in 9 provinces in China, including Beijing, Hebei, Jilin, Shandong, Shaanxi, Xinjiang, Anhui, Guangdong, Hunan were retrospectively analyzed for their respiratory samples, clinical and epidemic data.The respiratory samples were detected for HMPV by quantitative real-time PCR.Results:A total of 11 660 cases were tested for HMPV, involving 296 (2.54%) HMPV-positive cases.Among 296 HMPV-positive cases, 218 were single HMPV infection, and 78/296 (26.35%) were co-infected with one or more respiratory viruses.HMPV mainly affected children under 5 years of age (3.10%), and in this population, the proportion of pneumonia in HMPV co-infection cases was significantly higher than that of single HMPV infection.HMPV could be detected all year round, which was more popular in winter and spring, with the peak of HMPV epidemic in March.Conclusions:HMPV is one of the important pathogens causing acute respiratory infection in children, showing a clear seasonal epidemic.HMPV can be infected alone or in combination with other respiratory viruses, which may increase the risk of pneumonia in children.

Prevalencia del virus de la rinotraqueitis infecciosa bovina en el departamento del Caquetá, Amazonía Colombiana / Prevalence of bovine infectious rhinotracheitis virus in Caquetá department, Colombian Amazon

RESUMEN La rinotraqueitis infecciosa bovina (IBR) es una enfermedad ampliamente difundida en el mundo con gran repercusión socio-económica en las ganaderías bovinas. El objetivo del presente estudio fue determinar la prevalencia de IBR en 100 hatos del departamento del Caquetá, para lo cual, se muestrearon 960 bovinos mayores de 36 meses entre enero y marzo de 2016 en predios seleccionados a partir de los criterios: a) tamaño (50-180 hectáreas), b) con más de 10 vacas en ordeño, c) disponibilidad de los productores para cooperar y d) accesibilidad de las vías. Las muestras de suero sanguíneo se remitieron al Laboratorio de Diagnóstico Clínico Veterinario del ICA y se analizaron a través la prueba Elisa de bloqueo (BHV-1) gB. Los sueros con porcentaje de bloqueo superior al 55% se consideraron positivos a anticuerpos para IBR. Se encontró alta seroprevalencia (73,13%), mayor además en machos que en hembras (p < 0,05). A nivel municipal la seroprevalencia fue mayor en los municipios de El Doncello, Puerto Rico y San Vicente del Caguán y se encontró diferencia significativa (p < 0,05) entre los nueve municipios analizados. A nivel de hatos, la prevalencia fue del 99%. En conclusión, la prevalencia del virus de la rinotraqueitis infecciosa bovina (IBR) en bovinos de doble propósito del departamento del Caquetá fue muy alta.

ABSTRACT Infectious bovine rhinotracheitis (IBR) is a disease widely spread throughout the world with great socio-economic impact on bovine livestock. The objective of this study was to determine the prevalence of IBR in 100 herds in Caqueta department, for which 960 cattle older than 36 months between January and March 2016 were sampled in farms selected from the criteria: a) size (50-180 hectares), b) with more than 10 cows in milking, c) availability of producers to cooperate and d) accessibility of roads. Blood serum samples were sent to the ICA veterinary clinical diagnostic laboratory and analyzed using the Elisa blocking test (BHV-1) gB, where sera with blocking percentage greater than 55% were considered as positive for IBR antibodies. High seroprevalence was found (73.13%), being higher in males than in females (p < 0.05). At municipal level, seroprevalence was greater in El Doncello, Puerto Rico and San Vicente del Caguán municipalities, significant difference (p<0.05) was found between the nine municipalities analyzed. At herd level prevalence was of 99%. In conclusion the prevalence of Infectious Bovine Rhinotracheitis (IBR) virus in dual-purpose cattle of Caquetá department was very high.

Effect of simvastatin pretreatment on human metapneumovirus infection and its mechanism / 解放军医学杂志

Objective: To investigate the effect of simvastatin pretreatment on human metapneumovirus (hMPV) infection and the related mechanism. Methods: Human bronchial epithelial cells (16HBE) were used in vitro experiments and BALB/c mice were used in vivo experiments. They were randomly divided into control group, simvastatin group, hMPV group, hMPV+simvastatin group. After corresponding treatment, the fluorescence of hMPV was observed with microscopy; virus titer and the expressions of autophagy-related gene recombinant human autophagy-related protein 5 (ATG5), Beclin1, and microtubule-associated protein 1 light chain 3 (LC3) were detected by Fluorescence Quantitative Real-time PCR; Western blotting was performed to detect autophagy-related protein ATG5, Beclin1, LC3 and serine/threonine kinase/mammalian rapamycin target protein (AKT/mTOR) pathway proteins. Results: In vitro experiment, the direct and indirect fluorescence expressions of hMPV and the virus titer were lower in hMPV+simvastatin group (0.260 ± 0.018) than those in hMPV group (1.241 ± 0.030), while the mRNA expression levels of autophagy-related gene ATG5, Beclin1 and LC3 were higher in hMPV+simvastatin group [(7.31±0.15), (8.67±0.88) and (6.55±0.30), respectively] than those in control group [(1.00±0.06), (1.00±0.05) and (1.10±0.06), respectively]; the mRNA and protein levels of autophagy-genes were the highest in hMPV+simvastatin group and the differences were statistically significant (P<0.05). The AKT/mTOR pathway was inhibited in hMPV group and hMPV+simvastatin group. In vivo experiment, the virus titer of the lung tissue and of the lung tissue supernatant inoculated in Vero E6 were lower in hMPV+simvastatin group (0.75±0.26 and 0.42±0.17, respectively) than those in hMPV group (2.46±0.53 and 1.80±0.40, respectively). The mRNA and protein expression levels of autophagy-related genes ATG5, Beclin1 and LC3 were the highest in hMPV+simvastatin group (3.89±0.42, 3.13±0.26 and 3.56±0.22, respectively), higher than those in control group and hMPV group, the differences were statistically significant (P<0.05), and the AKT/mTOR pathway was inhibited in this group; HE staining of lung tissue showed an inflammatory response in the hMPV group, and pre-treatment with simvastatin could reduce the inflammation. Conclusion: Pretreatment with simvastatin may reduce human metapneumovirus infection, which is associated with autophagy and the AKT/mTOR pathway.

Protection against human metapneumovirus ( hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV / 中华微生物学和免疫学杂志

Objective To evaluate the immune responses and protection against human metapneu-movirus ( hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV. Methods Two recombinant influenza viruses ( rFLU/hMPV/B and rFLU/hMPV/CTL+Th ) carrying hMPV multi-epitope gene segments in NS gene were generated by reverse genetic techniques of eight-plasmid system. BALB/c mice were immunized intranasally with rFLU/hMPV/B and rFLU/hMPV/CTL+Th twice at a two-week interval. Virus-specific antibody titers and splenocyte cytokines were detected two weeks after the boost immunization. Viral loads in lung tissues and turbinates were detected with digital PCR after the immunized mice were challenged with hMPV and influenza virus. Moreover, HE staining was used to observe lung inju-ries. Results Specific antibodies against both the influenza virus and hMPV were induced in mice immu-nized intranasally with rFLU/hMPV/B, while the influenza virus-specific antibody response and hMPV-spe-cific cytotoxic lymphocyte response ( significant IFN-γ secretion ) were detected in mice immunized with rFLU/hMPV/CTL+Th. Additionally, balanced Th1/Th2 responses were elicited by rFLU/hMPV/B and rFLU/hMPV/CTL+Th. Both rFLU/hMPV/B and rFLU/hMPV/CTL+Th conveyed effective protection against subsequent influenza virus and hMPV challenges with significantly alleviated histopathological dama-ges and reduced viral loads. Conclusions Both rFLU/hMPV/B and rFLU/hMPV/CTL+Th can induce spe-cific humoral immune response against hMPV and/or the influenza virus. Moreover, rFLU/hMPV/CTL+Th can also elicit hMPV-specific CTL immune response. These two recombinant strains can also protect BALB/c mice from the challenges with hMPV and influenza virus, suggesting that they are promising vaccine candi-dates.

Protection against human metapneumovirus (hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV / 中华微生物学和免疫学杂志

Objective@#To evaluate the immune responses and protection against human metapneumovirus (hMPV) conveyed by influenza virus vectors carrying multiple epitope antigens of hMPV.@*Methods@#Two recombinant influenza viruses (rFLU/hMPV/B and rFLU/hMPV/CTL+ Th) carrying hMPV multi-epitope gene segments in NS gene were generated by reverse genetic techniques of eight-plasmid system. BALB/c mice were immunized intranasally with rFLU/hMPV/B and rFLU/hMPV/CTL+ Th twice at a two-week interval. Virus-specific antibody titers and splenocyte cytokines were detected two weeks after the boost immunization. Viral loads in lung tissues and turbinates were detected with digital PCR after the immunized mice were challenged with hMPV and influenza virus. Moreover, HE staining was used to observe lung injuries.@*Results@#Specific antibodies against both the influenza virus and hMPV were induced in mice immunized intranasally with rFLU/hMPV/B, while the influenza virus-specific antibody response and hMPV-specific cytotoxic lymphocyte response (significant IFN-γ secretion) were detected in mice immunized with rFLU/hMPV/CTL+ Th. Additionally, balanced Th1/Th2 responses were elicited by rFLU/hMPV/B and rFLU/hMPV/CTL+ Th. Both rFLU/hMPV/B and rFLU/hMPV/CTL+ Th conveyed effective protection against subsequent influenza virus and hMPV challenges with significantly alleviated histopathological damages and reduced viral loads.@*Conclusions@#Both rFLU/hMPV/B and rFLU/hMPV/CTL+ Th can induce specific humoral immune response against hMPV and/or the influenza virus. Moreover, rFLU/hMPV/CTL+ Th can also elicit hMPV-specific CTL immune response. These two recombinant strains can also protect BALB/c mice from the challenges with hMPV and influenza virus, suggesting that they are promising vaccine candidates.

Detection of avian metapneumovirus subtype A from wild birds in the State of São Paulo, Brazil / Detecção de metapneumovirus aviário subtipo A em aves silvestres no estado de São Paulo, Brasil

The present study investigated the circulation of avian metapneumovirus (aMPV) in wild birds in Brazil. To do so, 131 samples from 366 oropharyngeal or cloacal swabs collected from 18 species of birds were tested individually or in pools by RT-PCR. Samples detected by RT-PCR were selected for DNA sequencing. Thirteen (9.9%) samples were detected by the RT-PCR targeting the N gene and four out of 13 samples were sequenced. Sequencing results showed a high identity with the aMPV subtype A. Our results confirm the circulation of the aMPV subtype A in wild birds in Brazil even five years after its last detection.(AU)

O presente estudo investigou a circulação de metapneumovírus aviário em aves silvestres no Brasil. Para tanto, 131 amostras de 366 suabes orofaringeanos ou cloacais coletados de 18 espécies de aves foram testadas individualmente ou na forma de pools por RT-PCR. As amostras detectadas por RT-PCR foram selecionadas para sequenciamento. Treze (9,9%) das amostras foram detectadas por RT-PCR tendo o gene N como alvo; destas, quatro foram sequenciadas com sucesso. Resultados do sequenciamento mostraram alta identidade com o aMPV de subtipo A. Nossos resultados confirmam a circulação de aMPV subtipo A em aves silvestres no Brasil mesmo cinco anos após sua última detecção.(AU)

Construction of recombinant influenza virus carrying human metapneumovirus epitopes / 中华微生物学和免疫学杂志

Objective To construct and rescue recombinant influenza virus strains expressing hu-man metapneumovirus ( hMPV) epitopes. -ethods B cell, CTL and Th epitopes predicted by bioinformat-ics software were coupled together in different combinations. These different array genes were inserted into the NS1 gene of influenza virus strain A/PR/8/34 ( PR8 ) , respectively. Recombinant PR8 influenza virus vectors expressing different hMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric hMPV epitopes. Hemagglutination ( HA) titers, half tissue culture infection dose ( TCID50 ) and growth curves were detected. Results Interval sequences GPGPG and KK were introduced into hMPV epitope combinations to construct multi-epitope antigens (MEA). These MEA were inserted into the PR8 NS gene, respectively. Using 8 plasmid system, three recombinant influenza virus strains were rescued successfully. After cultured for three passages in Madin-Darby canine kidney ( MDCK) cells and one in eggs, these three recombinant strains could proliferate steadily. Whole genome sequencing verified that the three recombinant strains car-ried the chimeric MEA sequences, named as rFLU/hMPV/B, rFLU/hMPV/CTL-Th and rFLU/hMPV/B-Th. HA titers of the recombinant strains were 128, 128 and 256 using turkey erythrocyte, respectively. Their TCID50 were 107. 0/ml, 106. 8/ml and 107. 0/ml, respectively. Growth curve tests also verified that the recombinant strains could proliferate steadily in MDCK cells. Conclusions Three recombinant influenza vi-rus vector strains carrying the B cell, CTL and Th epitopes of hMPV were rescued successfully. This study lays the foundation for further evaluation of the immune effects of these recombinant viruses and their poten-tial application value in vaccine development.

Clinical characteristics and leukotriene levels in serum and urine of bronchiolitis children infected with different viruses / 中华实验和临床病毒学杂志

Objective@#To investigate the differences of clinical characteristics and leukotrienes (LTs) level of bronchiolitis children infected with respiratory syncytial virus (RSV), human rhinovirus (HRV) and human metapneumovirus (hMPV), and provide clinical evidence for the treatment of bronchiolitis with LTs receptor antagonist (LTRA) montelukast.@*Methods@#Totally 90 children with bronchiolitis hospitalized from January 2017 to December 2018 were enrolled into this study and viral nucleic acid from respiratory tract specimens were detected; and the patients were divided into three experimental groups: RSV group, HRV group and hMPV group. The clinical data and LTs level in blood and urine of experimental groups were compared; 30 healthy children were enrolled as the control group.@*Results@#There were no significant differences in age, sex, weight, family history and past history of allergy among the three experimental groups. The LTs levels in the experimental groups were higher than that of control group (P<0.05). There was a positive correlation between the LTs levels in blood and urine (r=0.723, P<0.05). In RSV group, LTs level of was the highest, lung function was the worst, and clinical score was the highest, and significantly different from those of HRV group and hMPV group (all P<0.01). However, there was no significant difference between HRV group and hMPV group. The number of days with fever of RSV group and hMPV group were not significantly different, but both were higher than that of HRV group (all P<0.01). The number of days with cough and wheezing among experimental groups had no significant difference(all P>0.05).@*Conclusions@#The bronchiolitis children infected with RSV had the highest LTs level, the worst lung function and the highest clinical score, there was significant difference when compared with HRV group and hMPV group.

Molecular prevalence and clinical characteristics of human metapneumovirus and human bocavirus in 333 hospitalized children with community-acquired pneumonia / 中华实验和临床病毒学杂志

Objective@#To study the molecular prevalence and clinical characteristics of human metapneumovirus and human bocavirus in hospitalized children with community-acquired pneumonia.@*Methods@#Total 333 throat swabs and clinical information of patients were collected between 2017 and 2018 at Department of Pediatrics of No.2 Clinical Teaching Hospital, Chengde Medical University. The IgM of adenovirus (AdV), respiratory syncytial virus (RSV), influenza virus-A/B (Influ-A/B), parainuenza viruses (PIVs)were tested by detection kit, and the positive samples of human metapneumovirus (hMPV), human bocavirus (HBoV), AdV, RSV and human coronavirus (HCoV) were detected by RT-PCR or PCR.@*Results@#43 cases, 19 cases, 3 cases and 2 cases were positive for Influ-B, PIV, RSV and AdV IgM, respectively. Total 80 cases were infected with hMPV (71 cases were single infection, 8 cases were double infection, and 1 case was triple infection), 22 cases were infected with HBoV (14 cases were single infection, 7 cases were double infection, and 1 case was triple infection), 6 cases were infected with AdV (4 cases were single infection, 1 case was double infection, and 1 case was triple infection), only 1 case was single infected with RSV or HCoV, respectively. 39 cases (11.7%) and 41 cases (12.3%) were distributed at <5 years group and ≥5 years group, respectively. 45.0%(18/40)in severe cases and 27.99%(82/293)in mild cases were positive for hMPV, HBoV, AdV, RSV and HCoV, the ratio of viral positive case was significant higher in severe cases than mild cases (P=0.042). The age (P=0.000), peak of fever (P=0.035), duration of hospitalization (P=0.000), neutrophils (P=0.000) and lymphocytes (P=0.000) were significant difference in severe cases compared with mild cases.@*Conclusions@#Multiple respiratory viruses can cause community-acquired pneumonia and more attention should be pay to the surveillance of hMPV and HBoV.

Viral etiology of acute respiratory infections in children in Southern Iran

Abstract INTRODUCTION: Prevalence of influenza A virus (Flu-A), respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) was assessed in children with acute respiratory infections (ARIs). METHODS: Nasopharyngeal aspirates and throat swabs were subjected to real-time polymerase chain reaction (PCR) to detect RSV and Flu-A and to conventional PCR to detect hMPV. RESULTS: Of the 156 children assessed, 93 (59.6%) carried at least one virus, with 35.9% positive for RSV, 14.1% for hMPV, and 9.6% for Flu-A. The prevalence of co-infections was 2.6%. CONCLUSIONS: The high detection rate may reflect increased sensitivity of real-time PCR compared to traditional PCR and viral culture.

Human metapneumovirus in Southern Brazil

INTRODUCTION Infections caused by respiratory viruses are important problems worldwide, especially in children. Human metapneumovirus (hMPV) is a respiratory pathogen and causes severe infections with nonspecific symptoms. This study reports the hMPV occurrence and dissemination in southern Brazil and compares the frequency of occurrence of this virus and the human respiratory syncytial virus (hRSV) in the epidemiological weeks in a three-year period (2009-2011). METHODS: In total, 545 nasopharyngeal (NP) specimens from individuals with Severe Acute Respiratory Syndrome (SARS) who were negative for other seven respiratory viruses were analyzed for the presence of hMPV. Human metapneumovirus was detected by direct immunofluorescence and real-time reverse transcription polymerase chain reaction. RESULTS: hMPV was detected in 109 patients from the main geographic regions of the southernmost state of Brazil, presenting similar overall prevalence in males (46.8%) and females (53.2%). Among children who were less than six years old, hMPV was detected in 99 samples of all age groups, with a higher frequency in infants who were less than one year old (45.7%) compared to all other age groups until six years. hMPV and hRSV infection occurred in almost the same epidemiological weeks (EWs) of each year, with peaks of incidence between EW 31/37 and EW 26/38 for the years 2009 and 2011, respectively. hMPV was further detected in several cases of SARS and it was the only virus detected in three deaths. CONCLUSIONS These findings indicate that hMPV is in circulation in southern Brazil and highlight the importance of diagnosing hMPV for influenza-like illness in the population. (AU)

Progress in detection methods of human metapneumovirus / 中华实验和临床病毒学杂志

Human metapneumovirus (hMPV) is a newly described paramyxovirus that was found in the Netherlands in 2001. It is an important pathogen of acute respiratory diseases in infants, young children, elderly and immunocompromised patients. hMPV infection could cause mild upper respiratory tract infection or severe lower respiratory tract disease, including bronchiolitis and pneumonia. hMPV was usually detected using direct immunofluorescence and RT-PCR, but the detection method were different according to the respective experiment requirements. In this paper we review the detection method of hMPV to provide a basis for further study of hMPV.

B1 subgenotype of human metapneumovirus detected in children with acute respiratory infection / 中华实验和临床病毒学杂志

Objective@#To reveal the prevalence of human metapneumovirus (HMPV) among pediatric patients with acute respiratory infection (ARI).@*Methods@#Nasopharyngeal aspirates were collected from hospitalized young children (1-5 years) diagnosed as ARI in Department of Respiratory Disease, Shanxi Provincial Children′s Hospital, Taiyuan city. The nucleoprotein (N) gene of HMPV was amplified by reverse transcription-polymerase chain reaction (RT-PCR). The positive products of PCR were sequenced, made homology and phylogenetic analysis.@*Results@#One sample collected in Feb. 2017 and two samples collected in Nov. 2017 were HMPV positive in RT-PCR detection. The homologies of nucleic acid and deduced amino acid sequences of the three positive PCR products with the corresponding HMPV sequences in GenBank were over 83% and 90%, respectively. Phylogenetic analysis revealed that HMPV B1 subgenotype was detected.@*Conclusions@#HMPV B1 subgenotype was the major epidemic HMPV in young children with ARI in Taiyuan city.

Comparison of nasal cytokine profiles of human metapneumovirus and respiratory syncytial virus

BACKGROUND: Human metapneumovirus (hMPV) and respiratory syncytial virus (RSV) share some epidemiological and clinical characteristics; however, few studies have examined the mechanisms by which these viruses induce airway inflammation. OBJECTIVE: This study was undertaken to compare cytokine profiles in hMPV and RSV patients to investigate possible differences in inflammatory pathways. METHODS: Nasopharyngeal aspirate specimens were collected from 1,008 pediatric patients hospitalized for acute lower respiratory tract infection with wheezing and 20 normal healthy controls. Patients were tested for 7 common respiratory viruses then divided into hMPV (n = 35) and RSV groups (n = 67). T helper (Th) 1 (interferon [IFN]-γ), Th2 (interleukin [IL]-4, eotaxin) and Th17 (IL-1β, IL-6) cytokine profiles were analyzed in the 3 groups. RESULTS: IFN-γ and IL-2 levels were significantly increased in the hMPV and RSV groups compared to the control group (p < 0.0001 and p < 0.0001, respectively). IL-4 levels were significantly higher in the RSV group compared to the hMPV and control groups (p = 0.0003 and p < 0.0001, respectively). Eotaxin levels showed a tendency to be higher in the RSV group compared to the hMPV group (p = 0.0580), and significantly higher compared to the control group (p < 0.0001). IL-1β levels were significantly higher in the hMPV compared to the RSV group (p < 0.0001), and IL-6 levels were significantly higher in the hMPV group compared to the control group (p < 0.0001). CONCLUSION: Our results suggest that hMPV and RSV have different inflammatory mechanisms. hMPV induces airway inflammation by the Th17 pathway through release of IL-1β and IL-6, whereas RSV acts through the Th2 pathway.

Influence of coinfection factors on clinical features of human metapneumovirus infection / 中华实用儿科临床杂志

Objective To explore the influence of coinfection with other pathogens on human metapneumovirus (hMPV) infection in children.Methods A total of 11 299 children admitted to the Department of Respiratory Disease,Children's Hospital of Soochow University between June 2010 and May 2015 were enrolled in this study.Sputum specimens were collected and multiple pathogenic joint detection was done,including peripheral blood,and blood routine,C reactive protein (CRP),hepatic function and cellular immunity.Patients' clinical data were collected.Results Among 11 299 hospitalized children,hMPV was positive found in 222 children (1.96％).One hundred and fourteen children (51.4％) had hMPV simple infection and 108 cases of them (48.6％) were coinfected with other pathogens.The hMPV coinfected with bacteria (63 cases,28.4％)was most common.The average age of multiple coinfected children was older than that of simple hMPV infection in children [(2.43 ± 2.47) years old vs.(1.27 ± 1.30) years old],and the difference was significant (Z =-2.360,P ＜ 0.05).Fever seemed to be more common in children coinfected with bacteria or multiple coinfection (63.5％ and 70.0％) compared with those with simple hMPV infection (43.0％),and the differences were significant (x2 =6.827,4.986,all P ＜ 0.05).There was no significant difference in other clinical features among 5 groups (all P ＞ 0.05).Multiple coinfection children had a higher percentage of neutrophils (0.50 ± 0.18) than that in simple hMPV infection children (0.37 ± 0.19),children coinfected with bacteria (0.39 ±0.19) or other virus (0.35 ±0.19),and the differences were significant (all P ＜0.05).CRP was elevated in 30.2％ (19/63 cases) of children coinfected with bacteria,which was significantly higher than that of simple hPMV infection children (16.7 ％,19/114 cases),and the difference was significant (x2 =4.381,P ＜ 0.05).CD3 + CD4 + was significantly lower in multiple coinfection children (0.31 ± 0.07),but there were no significant difference compared with other groups (all P ＞ 0.05).CD19 + CD23 + was significantly higher in children coinfected with other virus com pared with that of simple hMPV infection group,hMPV coinfected with bacteria,hMPV coinfected with Mycoplasma pneumonia and multiple coinfect group (0.37 ± 0.10 vs.0.30 ± 0.09,0.30 ± 0.08,0.29 ± 0.07,0.29 ± 0.09),and the differences were significant (all P ＜ 0.05).Conclusions It is suggested that hMPV seems easily coinfected with other pathogens,especially with bacteria.It should be on high alert that bacteria coinfection is accompanied with high percentage of neutrophils and high level of CRP.Coinfection does not significantly exacerbate the clinical symptoms of hMPV infection,but may exacerbate the cellular immune disorders to a certain extent.

Study on Causes of Respiratory Disease to the Middle East Respiratory Syndrome-negative Subjects

Middle East respiratory syndrome coronavirus (MERS-CoV) causes severe cases of human respiratory disease. The current outbreak of infection with this virus in South Korea, which began on May 20, 2015, has infected 186 patients and caused 36 deaths within 2 months. In this study, to investigate the viral pathogen causing acute respiratory infections, multiplex/RT-PCR was performed on were obtained from nucleic acid of the Middle East Respiratory Syndrome-negative subjects. Viruses and atypical bacteria were detected in 39 of 337 (11.6%). Frequent viruses were human rhinovirus (n=11, 3.3%), human metapneumovirus (n=9, 2.7%), parainfluenza (n=9, 2.7%) and adenovirus (n=4, 1.2%). Mycoplasma pneumonia (M. pneumonia) was detected in 1.8 % (n=6). Out of 9 human metapneumovirus (hMPV) positive samples, 6 samples were successfully sequenced using F gene. And M. pneumoniae was sequencing of a repetitive region of the P1 gene. Phylogenetic analysis revealed that hMPV clustered into A2b lineage (n=4), B2 lineage (n=2) and M. pneumoniae clustered into two genotypes: Type 1 (n=4), Type 2a (n=2).