我国急性呼吸道感染患儿中检测到KI和WU多瘤病毒

目的 了解多瘤病毒WU和KI在我国儿童急性呼吸道疾病中的感染情况.方法 采用PCR扩增的方法对2006年11月至2007年10月收集的急性呼吸道感染患儿的318份鼻咽抽吸物(NPA)标本进行了多瘤病毒WU和KI基因检测.结果 318份标本共检测出14份病毒核酸阳性标本,其中WUV 7份(2.2%),KIPyV 7份(2.2%).该14例基因检测阳性患儿临床均有上呼吸道感染或下呼吸道感染症状.结论 WUV和KIPyV可能也是儿童急性呼吸道感染中较为重要的一个病原,且与儿童上呼吸道感染和下呼吸道感染存在相关性.     

WU多瘤病毒-新发现的人类多瘤病毒的研究概况

随着人类多瘤病毒的不断被发现,人们对它的研究兴趣也越来越大.自2007年人类WU多瘤病毒(WU polyomavirus,WUPyV)在呼吸道标本中被发现以来,世界各地陆续发表了发现WUPyV的报道.WUPyV在全世界广泛流行,可能在儿童时期获得感染,但大多数不表现临床症状.尽管WUPyV经常在呼吸道标本中发现,但是WUPyV是否为呼吸道疾病的病原还有争议.WUPyV的传播途径和流行性也有待进一步的研究.目前该病毒的流行病学资料主要来自PCR检测和血清学试验.本文现就WU多瘤病毒的研究进展作一综述.     

WU多瘤病毒-新发现的人类多瘤病毒的研究概况

随着人类多瘤病毒的不断被发现,人们对它的研究兴趣也越来越大.自2007年人类WU多瘤病毒(WU polyomavirus,WUPyV)在呼吸道标本中被发现以来,世界各地陆续发表了发现WUPyV的报道.WUPyV在全世界广泛流行,可能在儿童时期获得感染,但大多数不表现临床症状.尽管WUPyV经常在呼吸道标本中发现,但是WUPyV是否为呼吸道疾病的病原还有争议.WUPyV的传播途径和流行性也有待进一步的研究.目前该病毒的流行病学资料主要来自PCR检测和血清学试验.本文现就WU多瘤病毒的研究进展作一综述.     

WU多瘤病毒-新发现的人类多瘤病毒的研究概况

随着人类多瘤病毒的不断被发现,人们对它的研究兴趣也越来越大.自2007年人类WU多瘤病毒(WU polyomavirus,WUPyV)在呼吸道标本中被发现以来,世界各地陆续发表了发现WUPyV的报道.WUPyV在全世界广泛流行,可能在儿童时期获得感染,但大多数不表现临床症状.尽管WUPyV经常在呼吸道标本中发现,但是WUPyV是否为呼吸道疾病的病原还有争议.WUPyV的传播途径和流行性也有待进一步的研究.目前该病毒的流行病学资料主要来自PCR检测和血清学试验.本文现就WU多瘤病毒的研究进展作一综述.     

长春地区2010年度婴幼儿腹泻的病原学研究及流行病学特点

目的 了解长春地区四种主要腹泻病毒病原构成及流行病学特点.方法 收集长春市儿童医院5岁以下住院患儿腹泻样本共460例,轮状病毒采用ELISA试剂盒检测,杯状病毒、星状病毒采用逆转录-聚合酶链反应( RT-PCR)法,腺病毒采用聚合酶链反应(PCR)法进行鉴定.结果 460份标本中轮状病毒占35.22％( 162/460);杯状病毒占20.43％( 94/460),星状病毒占9.78％(45/460),腺病毒占3.70％(17/460),混合感染达7.17％(33/460),发病患儿以2岁以下婴幼儿为主.对162份轮状病毒阳性标本进行G/P分型,结果示G1P[8]为主要流行株,杯状病毒以GⅡ-4亚型为主要流行株,星状病毒为Ⅰ型,腺病毒为Ad41.结论 长春地区婴幼儿病毒性腹泻的病原中轮状病毒是最主要病原,其次为杯状病毒、星状病毒和腺病毒.     

深圳市某福利院一起急性结膜炎疫情的病原学分析

目的 鉴定深圳市某福利院一起急性结膜炎疫情的病原体.方法 采用荧光定量PCR方法对12份眼拭子标本进行检测以确定病原,阳性标本进行病毒分离培养,扩增特异基因并测序,对序列进行进化分析和分子分型.结果 12份标本荧光定量PCR检测结果为人腺病毒阳性,分离9株病毒.人腺病毒Hexon基因测序显示9株腺病毒之间核苷酸相似性在98.3％-100％之间,系统进化分析显示9株腺病毒与人腺病毒7型参考株位于同一进化分支.结论 本起疫情由人腺病毒7型引起,与我国近期流行的腺病毒7型同源.     

太原市5岁以下腹泻住院儿童的病毒病原及其流行特征

目的 了解山西省太原市5岁以下腹泻住院儿童四种主要腹泻病毒的流行情况.方法 收集山西儿童医院2007年10月至2008年11月5岁以下全部住院腹泻患儿的粪便标本,采用ELISA试剂盒来检测轮状病毒;采用聚合酶链反应（PCR）检测腺病毒;逆转录-聚合酶链反应（RT-PCR）法检测星状病毒和杯状病毒并对轮状病毒进行分型.结果 346份标本中轮状病毒占40.8%、杯状病毒占7.5%、星状病毒占6.4%、腺病毒占3.2%.对141份轮状病毒阳性标本进行G1P分型,G1型是最优势株,P型优势株为P[8].四种病毒主要是感染2岁以下婴幼儿,RV有明显的季节的特征,9-11月份（48.92%）为发病高峰.结论 轮状病毒为最主要的病毒病原,G1P[8]型为主要流行株,秋冬季为发病高峰,2岁以下为发病高危人群,混合感染多见.     

不同方法检测两种呼吸道病毒结果的比较

呼吸道合胞病毒 (ｒｅｓｐｉｒａｔｏｒｙｓｙｎｃｙｔｉａｌｖｉｒｕｓ,ＲＳＶ)、腺病毒(ａｄｅｎｏｖｉｒｕｓ ,ＡＤＶ)是引起儿科急性下呼吸道感染的两种重要病原。实验室提供及时和准确的病原检测报告 ,对临床明确病因、采取适当治疗手段和防止滥用抗生素有重要意义。自 90年代起 ,国外有报道将快速细胞培养和免疫组化的方法结合在一起检测病毒。本研究采用美国Ｃｈｅｍｉｃｏｎ公司生产并由ＷＨＯ认可的免疫荧光试剂 ,用直接涂片和快速细胞培养法分别检测呼吸道合胞病毒和腺病毒 ,并同时做常规的病毒分离 ,目的在于比较和评价不同方法…     

一种检测腹泻相关病毒的方法

目的 建立检测腹泻粪便标本中病毒病原的方法.方法 选取本实验室检测的杯状病毒、肠道腺病毒、轮状病毒、博卡病毒、星状病毒以及肠道病毒阳性标本各一份,通过非序列依赖性单-PCR扩增(Sequence-independent single primer amplification,SISPA)构建基因文库,从中筛查病毒基因片段.结果 六份标本中都可以得到相对应的病毒核酸序列.结论 本研究采用的方法可以检测到粪便标本中引起腹泻的相关病毒,为进一步研究检测目前尚未发现的病毒病原奠定基础.     

肾移植后多瘤病毒相关性肾病

背景：多瘤病毒感染是导致多瘤病毒相关性肾病和移植肾失功的重要原因之一。 目的：观察分析多瘤病毒相关性肾病的临床特征及其病理学特点。 方法：121例患者移植肾活检行多瘤病毒大T抗原染色,发现9例阳性诊断为多瘤病毒相关性肾病,利用SV-40 大 T 抗原免疫组织化学染色,对确认为多瘤病毒相关性肾病患者进行临床、病理、免疫荧光、免疫组织化学观察。 结果与结论：多瘤病毒相关性肾病组肾活检时检测霉酚酸-AUC 0-12和他克莫司血药浓度均明显高于同期非多瘤病毒相关性肾病组(P < 0.05)。9例活检组织经SV-40 大T抗原染色,肾皮质和髓质均可见散在的肾小管多瘤病毒阳性。免疫荧光IgG,IgM,IgA,C3,C4,C1q和C4d全阴性,所有肾组织病理均可见肾间质大量聚集的CD3,CD4,CD8,CD68阳性细胞,1例合并排斥反应者,人白细胞DR抗原和白细胞介素2受体高表达;不合并排斥者人白细胞DR抗原和白细胞介素2受体表达多小于5%。9例多瘤病毒相关性肾病患者随访均超过半年,移植肾失功1例,3例好转,2例稳定,3例恶化。结果表明,多瘤病毒相关性肾病的诊断主要依赖于移植肾活检组织病理;利用SV-40 大T 抗原免疫组织化学染色可提高多瘤病毒相关性肾病的诊断率;移植肾组织C4d、白细胞介素2受体和人白细胞DR抗原检测对多瘤病毒感染相关性肾病的鉴别诊断具有极其重要的临床价值。     

Diagnosis of psittacine beak and feather disease (PBFD) viral infection, avian polyomavirus infection, adenovirus infection and herpesvirus infection in psittacine tissues using DNA in situ hybridization

The evaluation of the usefulness of DNA probes in a diagnostic setting to identify nuclear inclusions in selected viral infections (psittacine beak and feather disease viral infection, avian polyomavirus infection, adenovirus infection and Pacheco's parrot disease) is reported. A DNA in situ hybridization method was used to detect viral nucleic acid in sections of paraffin-embedded tissues coming from birds naturally and/or experimentally infected. It is concluded that DNA probes used for polyomavirus (FN-19) and adenovirus (FN-23) are able to identify nucleic acid of each virus in the cells with nuclear inclusions, and when used for psittacine beak and feather disease virus (FN-8), and Pacheco's parrot disease virus (FN-49) are able to detect viral nucleic acid in cells with or without inclusions.     

Prevalence and genetic characterization of avian polyomavirus and psittacine beak and feather disease virus isolated from budgerigars in Mainland China

Budgerigar fledgling disease (BFD) and psittacine beak and feather disease (PBFD) are caused by avian polyomavirus (APV) and psittacine beak and feather disease virus (PBFDV), respectively. These diseases frequently infect psittacine birds and result in similar clinical manifestations. In this study, we observed the prevalence of PBFDV infection and a dual infection of APV and PBFDV in a budgerigar (Melopsittacus undulatus) in Mainland China for the first time. One PBFDV isolate and two APV isolates were harvested using chicken embryos. Genetic characterization and phylogenetic analysis of the complete genome of the two APV isolates revealed nucleotide similarity ranging from 99.0% to 99.6% to other sequences in GenBank, and a 14-bp insertion was observed in the genome of one APV isolate. The results of complete genome analysis of the PBFDV isolate showed nucleotide similarity ranging from 83.0% to 95.0% with other PBFDV sequences in GenBank. Genetic characterization and phylogenetic analysis of the APV and PBFDV strains isolated in this study indicated that the isolates from China were closely related to their Japanese counterparts. The results of this study will help to identify molecular determinants and will aid further research on the prevention and control of APV and PBFD infection.     

Nucleotide sequence analysis of a C1 gene fragment of psittacine beak and feather disease virus amplified by real-time polymerase chain reaction indicates a possible existence of genotypes

To investigate sequence diversity of psittacine beak and feather disease virus, samples collected from 31 psittacine species with or without clinical signs were tested for the presence of the viral genome. A real-time polymerase chain reaction was developed amplifying a 202 base pair fragment of the region encoding the capsid protein C1 and detecting 100 to 1000 genome equivalents. The nucleotide sequences of the polymerase chain reaction products showed 84.1 to 100% identity with no consistent pattern with regard to the infected bird species. Amino acid exchanges were concentrated mainly in five of the 42 deduced positions. Sequences obtained from an outbreak of acute beak and feather disease in lories clustered in a separate branch of a phylogenetic tree. Sequences in samples from African grey parrots with feather disorders grouped together, whereas those from the same species with immunosuppression clustered in other branches. These results indicate the possible existence of beak and feather disease virus genotypes.     

Comparitive sensitivity of polymerase chain reaction diagnosis of psittacine beak and feather disease on feather samples, cloacal swabs and blood from budgerigars (Melopsittacus undulates, Shaw 18005).

A longitudinal study was performed in order to investigate virus excretion and viraemia during a clinical outbreak of the psittacine beak and feather disease in budgerigars (Melopsittacus undulatus). Viral nucleic acid was detected in feathers, cloacal swabs and blood samples. Overall, beak and feather disease virus (BFDV) DNA was detected most commonly in feather samples, followed by cloacal swabs, and least frequently from blood samples. In most cases the viraemia was short lived and correlated with clinical signs, such as feather abnormalities. Sequence analysis of the polymerase chain reaction fragment amplified from the replication-associated gene (ORF V1) indicated a close relationship with other BFDV isolates. Overall the highest level of nucleotide identity was found with the ORF V1 of another budgerigar isolate. Our results suggest that feather samples and cloacal swabs should be taken for polymerase chain reaction diagnosis to determine the presence of BFDV in an aviary, but that detection in these samples may not correlate well with psittacine beak and feather disease.     

Development of novel real-time PCR assays for detecting DNA virus infections in psittaciform birds

Viral diseases of psittacine birds are detected presently by PCR. However, conventional PCR methods are not quantitative and the products can sometimes include non-specific products of the same size. To avoid these problems, real-time PCR assays based on the SYBR Green assay system were developed for the detection and quantitation of four virus diseases of psittacine birds: psittacine beak and feather disease, avian polyomavirus infection, psittacid herpesvirus infection, and psittacine adenovirus infection. Up to 1x10(2) copies of virus DNA were detected, indicating that these assays are as sensitive as conventional PCR assays. The assays are specific because they did not amplify any other pathogens including other viruses, bacteria, and fungi in psittacine birds. The assays measured successfully virus loads in clinical samples (blood, feathers, and tissues), showing that these specimens were suitable targets for the detection and quantitation of viral DNA in psittacine birds.     

Beak and feather disease virus infection in cockatiels (Nymphicus hollandicus).

Psittacine beak and feather disease is known to occur in a wide range of psittacine species; however, there are no scientific or credible anecdotal reports of psittacine beak and feather disease occurring in the cockatiel (Nymphicus hollandicus) despite it being one of the world's most commonly kept companion bird species. Consequently, this has resulted in speculation that the species may have some innate resistance to beak and feather disease virus (BFDV) infection. To investigate this hypothesis we conducted a survey of cockatiels (n=88) at commercial aviaries to investigate whether BFDV infection occurs in cockatiels, and found that all birds were virus-free by polymerase chain reaction and haemagglutination assay and had no detectable antibody titre by haemagglutination-inhibition assay. In addition to this, we sequenced the genome of two BFDV isolates obtained from diseased cockatiel feathers and performed cross-reactivity assays using virus eluted from these feathers and sera from naturally immune psittacine birds. Serological cross-reactivity results and phylogenetic analysis of the nucleotide sequences indicated that the cockatiel virus isolates were serologically and genetically different to other BFDV isolates. This is the first paper to report evidence of an antigenically distinct BFDV in psittacine birds.     

Hematology of vaccinated and non-vaccinated long-billed corellas following infection with beak and feather disease virus (BFDV)

The hematological characteristics of juvenile long-billed corellas (Cacatua tenurostris), with or without prior administration of a psittacine beak and feather disease vaccine, were studied for 97 days after experimental infection with beak and feather disease virus (BFDV). It was found that the pre-challenge hematological values were similar between vaccinated and non-vaccinated corellas. Most pre-challenge parameters were comparable to previously reported values of other cockatoos and psittacine birds. Significant differences were seen in both groups when comparing pre-challenge values with post-challenge values for total and differential leukocyte concentrations, but packed cell volume and total serum protein were not significantly affected by BFDV challenge.     

Molecular analysis and associated pathology of beak and feather disease virus isolated in Italy from young Congo African grey parrots (Psittacus erithacus) with an “atypical peracute form” of the disease

This study is the first report on the genetic and pathogenic characterization of beak and feather disease virus (BFDV) occurring in Italy. Twenty BFDV strains isolated in Italy from juvenile Congo African grey parrots (Psittacus erithacus) were investigated. Seventeen strains showed an “atypical peracute form” (aPF) of the disease, and three a chronic form (CF). The birds with aPF had been weaned, were independent as far as food and protection were concerned and apparently were without lesions. The gene coding for the putative coat protein was amplified in all isolates while the BFDV genome was sequenced completely in 10 samples, eight of them belonging to aPF affected birds and two from CF of the disease. All full genomes clustered into the J strain of BFDV, where two new subtypes were identified. Recombination analyses showed evidence of genetic exchanges in two BFDV genomes. In addition, a correlation between viral isolate and origin of the breeding material was shown, while an association between the genetic features of the virus and the clinical form was not observed. Histologically, apoptosis was detected frequently in aPF samples and sporadically in CF samples. Interestingly, BFDV antigens were detected in the nuclei and cytoplasm of such apoptotic cells. The data presented here support the hypothesis that, in the absence of a defined BFDV genetic variant accountable for a specific clinical form of psittacine beak and feather disease, differences in the apoptotic rate between aPF and CF are strictly host related.     

Reovirus infections associated with high mortality in psittaciformes in The Netherlands.

In The Netherlands between January 2002 and December 2004, numerous psittaciformes died showing severe splenomegaly and hepatomegaly with multifocal acute necrosis. At the start of the outbreaks mostly parakeets were affected, but later larger parrots were also involved. Seventy-eight birds showed the same features and six were examined completely, including a virological examination. Tests for polyomavirus, Pacheco's disease (herpesvirus) and circovirus psittacine beak and feather disease (PBFD) viruses and Chlamydophila psittaci were carried out. All results were negative, except for two cases of circovirus infection. Many concurrent bacterial and parasitic infections were seen. Immunohistochemistry revealed reovirus antigen in intralesional mononuclear cells, and reovirus-like particles could be observed by negative contrast electron microscopy. A reovirus was grown and the isolates reacted with polyclonal reovirus antiserum but did not react with monoclonal antibodies against chicken reovirus. The virus was therefore considered a psittacine reovirus. Because reoviruses were seen consistently, they seemed to be the most probable cause of the outbreaks. Climate, the introduction of new birds and the transportation of birds might be other factors involved in the disease seen in The Netherlands. No regional influence could be seen; therefore, we suggested that the virus might be widespread and carriers could be a source of re-introduction.