Animals ; Humans ; Virus Diseases ; virology ; Viruses ; classification ; genetics ; isolation & purification

In nature, honeybees are the most important pollinators. They play a vital role in both protecting the diversity of natural ecosystems, and maintaining the yield-improving effects of agroecosystems. But in recent years, epidemic disease in bees has caused huge losses. Black Queen Cell Virus (BQCV) is a bee pathogen that was first reported in 1955. It mainly infects bee larvae and pupae, making their bodies turn dark and black, and causing a massive decrease in the bee population. More specifically, the virus makes the exterior of the cell walls in the larvae and pupae turn black. BQCV is a seasonal epidemic, spread by means horizontal and vertical transmission, and is often unapparent. BQCV not only infects a variety of bee species, but also spiders, centipedes and other arthropods. It can also be coinfected with other honeybee viruses. In recent years, research has shown that the Nosema intestinal parasite plays an important role in BQCV transmission and bees carrying Nosema that become infected with BQCV have increased mortality. Here we summarize current research on the incidence, prevalence, geographical distribution and transmission of BQCV.
Animals ; Bees ; virology ; Dicistroviridae ; classification ; genetics ; isolation & purification ; physiology ; Insect Viruses ; classification ; genetics ; isolation & purification ; physiology

OBJECTIVE: Tick-borne encephalitis virus (TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis (TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. METHODS: A reverse-transcription recombinase-aided amplification (RT-RAA) assay was developed. This assay can be completed in one closed tube at 39 °C within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type (WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. RESULTS: The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units (pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. CONCLUSION A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.
Encephalitis Viruses, Tick-Borne ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; RNA, Viral ; analysis

OBJECTIVETo investigated the molecular epidemiologic features of viral diarrhea in Chengdu infants and young children, and to establish baseline patterns of etiology, provides the scientific basis for the vaccine development and the epidemic situation control.

METHODSFrom March, 2006 to December, 2008, a total of 376 infants and young children from Chengdu area hospitalized for diarrhea in Chengdu Children's Hospital were enrolled in this study. The stool specimen collected from each patient was tested for rotavirus (RV), Calicivirus (CV), astrovirus (AstV) and adenovirus (Adv) by using enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) examination.

RESULTSAmong those 376 cases,there were 142 cases (37.76%) of RV infections,which scattered predominantly in October to December. Among 234 cases RV negativity,there were 29 cases HuCV infections (15.85%), 5 cases AstV infections (1.64%), and 8 cases Adv infections (2.04%).

CONCLUSIONRV appeared to be the main etiological agent of viral diarrhea in Chengdu infants and young children,the predominant serotype of RV were G3, P[8] and P[4],HuCV might be the important etiological agent besides RV.

Adenoviruses, Human ; genetics ; isolation & purification ; Caliciviridae ; genetics ; isolation & purification ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Mamastrovirus ; genetics ; isolation & purification ; Molecular Epidemiology ; Rotavirus ; genetics ; isolation & purification ; Virus Diseases ; epidemiology ; virology ; Viruses ; genetics

OBJECTIVETo study viruses infecting Pinellia ternata in China.

METHODSymptom observation, DAS-ELISA and RT-PCR detection were applied.

RESULT AND CONCLUSIONDuring a survey in early spring, SMV and CMV were both commonly distributed as main viruses infecting P. ternata collected from different areas in China. But DsMV was the virus which infected P. ternate in natural condition. The infection ratio of cultivated P. ternate by SMV and CMV were 71.4% and 14.3% respectively for 21 samples collected from Ningbo, Zhejiang province; 100% and 44.4% for 18 samples from Xiaoshan, Zhejiang province; 61.9% and 33.3% for 21 samples from Hebei province; 50.0% and 41.7% for 12 samples from Anhui province; 16.7% and 16.7% for 12 samples from Sichuan province; 31.3% and none for 16 samples from Beijing. And the infection ratio of 25 wild samples from different areas of China infected by SMV and CMV were both 20.0%.

China ; Cluster Analysis ; Cucumovirus ; genetics ; isolation & purification ; DNA, Complementary ; chemistry ; genetics ; Mosaic Viruses ; classification ; genetics ; isolation & purification ; Pinellia ; virology ; Plant Diseases ; virology ; Plants, Medicinal ; virology ; Sequence Analysis, DNA

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.
Arboviruses ; genetics ; isolation & purification ; Encephalitis Virus, Japanese ; genetics ; Encephalitis Viruses ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

PURPOSE: Early identification of causative agents in lower respiratory infection of pediatric patients can reduce morbidity and prevent an overuse of antimicrobials. Two kinds of multiplex polymerase chain reaction (PCR) and a commercial shell vial viral culture were performed to identify causative agents in pediatric patients. MATERIALS AND METHODS: Nasopharyngeal aspirates of 220 children diagnosed with viral pneumonia were obtained. Two kinds of multiplex PCR (Seeplextrade mark RV detection kit, and Labopasstrade mark RV detection kit), and a shell vial culture by R-Mix were performed. RESULTS: Positive samples from 220 total samples by two multiplex PCRs were 52.7% and 46.4%, respectively. We also cultured 103 samples that showed positive results of the adenovirus, influenza virus, parainfluenza virus, and respiratory syncytial virus (RSV) by two multiplex PCR. The RSV was most frequently detected in 53.0% (Seeplex) and 51.7% (Labopass) of patients. The detection rate of adenovirus (AdV) was 10.3% and 12.1%, influenza virus (IFV) A and B was 12.5% and 3.4%, and parainfluenza virus (PIFV) 1, 2, and 3 were 2.9% and 2.6%. Shell vial cultures showed concordant results with each multiplex PCR by 96.1% and 77.7%, respectively. Sequencing results were 90% consistent with multiplex PCR. CONCLUSION: Multiplex PCR showed more positivity than the shell vial culture and it can be an effective primary test. Other complementary efforts such as viral cultures and sequencing analysis could be considered, according to clinical and laboratory conditions.
Adenoviridae/genetics/*isolation & purification ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Orthomyxoviridae/genetics/*isolation & purification ; Pneumonia, Viral/*virology ; Polymerase Chain Reaction/*methods ; Respiratory Syncytial Viruses/genetics/*isolation & purification ; Respirovirus/genetics/*isolation & purification

A GeXP based multiplex RT-PCR assay was developed to simultaneously detect twelve different respiratory viruses types/subtypes including influenza A virus, influenza B virus, influenza A virus sH1N1, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3, human rhinovirus, human metapneumovirus, adenovirus, respiratory syncytial virus A, respiratory syncytial virus B and human bocavirus. Twelve sets of specific primers were designed based on the conserved sequences of available respiratory-virus sequence database. The specificity of the multiplex system was examined by positive specimens confirmed previously. The sensitivity to detect twelve respiratory viruses simultaneously was 10(3) copies/microL. Twenty four clinical specimens were further detected by this novel assay and the results were compared with that of the real-time RT-PCR. These results showed that this novel assay based on GeXP is a fast, sensitive, and high throughput test for the detection of respiratory virus infections.
Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Orthomyxoviridae ; genetics ; isolation & purification ; Orthomyxoviridae Infections ; virology ; RNA Viruses ; genetics ; isolation & purification ; Real-Time Polymerase Chain Reaction ; instrumentation ; methods ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods ; Rhinovirus ; genetics ; isolation & purification ; Sensitivity and Specificity

Genetic Techniques ; Genomics ; Humans ; Nucleic Acid Amplification Techniques ; Nucleic Acid Hybridization ; Sequence Analysis, DNA ; Viruses ; genetics ; isolation & purification

Viral hemorrhagic fevers (VHFs) refer to a group of acute infections with high case fatality rates that are caused by four distinct families of RNA viruses belonging to the families Bunyaviridae, Flaviviridae, Filoviridae and Arenaviridae, the main clinical symptoms of these diseases are accompanied by fever and bleeding. For the reason that these infections have similar primary clinical symptoms, it is difficult to diagnose and distinguish them; rapid and reliable laboratory diagnostic tests are required in suspected cases for epidemiological investigation and controlling the spread of VHFs. This review addresses the laboratory diagnostics of VHFs, covering etiological classification and different diagnostic techniques, such as virus isolation, nucleic acid detection, as well as antigen and antibody assays. Prospects for novel diagnostic tools are also discussed.
Clinical Laboratory Techniques ; methods ; Hemorrhagic Fevers, Viral ; diagnosis ; immunology ; virology ; Humans ; RNA Viruses ; genetics ; immunology ; isolation & purification