Genome amplification and sequence analysis of human bocavirus 2 / 病毒学报

To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.

Molecular and epidemiological study on among children under 5 years old in Nanjing / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the infected information, clinical symptom and molecular epidemiological characteristics of HuCV infection among children under 5 years old in Nanjing.</p><p><b>METHODS</b>In Nanjing Children's Hospital of Nanjing Medical University from July 2010 to June 2011, we collected 428 stool specimens from children with diarrhea and 428 asymptomatic controls. Human Calicivirus were tested by using RT-PCR. Then we sequenced the nucleic acid of PCR amplifications and identified the genotype and gene group of prevalent strains.</p><p><b>RESULTS</b>63 (14.72%) out of 428 stool samples were detected as HuCV. 58 were norovirus and 5 were sapovirus, while GII-4 2006b was the predominant strain of NoV. In the 428 control samples, 19 samples were positive for calicivirus, there were 8 NoV and 13 SaV (Including 3 co-infection cases).</p><p><b>CONCLUSION</b>Human caliciviruses with different genotypes circulated among children in Nanjing,and GII. 2006b is the dominant genotype.</p>

A novel method for multiplex detection of gastroenteritis-associated viruses / 病毒学报

To develop and optimize a simultaneous detection method of RotavirusA, Norovirus GI, GII, Sapovirus, human astrovirus, enteric adenoviruses and HBoV2 with GenomeLab GeXP analysis system. The sensitivity was verified to be 10(4) copies/microL with plasmids containing the viral targets in triplicate on different days, and no cross-reaction with enterovirus71, human Parechovirus and PicobirnavirusII was observed. Finally, we successfully developed a high throughout, rapid and maneuverable multiplex RT-PCR assay for simultaneous detection of seven viruses related with viral gastroenteritis, which provide a novel method for the molecular diagnosis of diarrhea-associated virus.

Molecular and epidemiological study on viral diarrhea among infants in Lanzhou / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the epidemiologic characteristics of viral diarrhea in children under 5 years old in Lanzhou, understand the four major virus in children of distribution.</p><p><b>METHODS</b>In the first hospital of Lanzhou university from Jul 2009 to Jun 2010,we collected 290 stool specimens from children with diarrhea and 114 asymptomatic controls. Rotavirus was detected by ELISA,further strain characterization was carried out by nested PCR. The human calicivirus, astrovirus, adenovirus were detected by RT-multiplex PCR and PCR.</p><p><b>RESULTS</b>At least one of the four viral agents was found in 60% of the specimens. Rotavirus, human calicivirus, adenovirus, and astrovirus were identified in 39.31%, 11.38%, 10.69%, and 4.83% in 290 specimens respectively. Rotavirus G3 was the most prevailing serotype, P [8] was the most common genotype. In the 114 control samples, 7 sample was positived for calicivirus, 5 samples were positived for human adenovirus and 1 sample was positived for astrovirus.</p><p><b>CONCLUSION</b>The results indicated clearly the impact of viral agents causing diarrhea and the importance of long-term systematic surveillance.</p>

Detection and molecular characterization of human parechovirus (HPeV) in children with acute gastroenteritis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study HPeV from stool samples of children with acute gastroenteritis under 5 years old.</p><p><b>METHODS</b>We conducted a real-time PCR to detect HPeV from stool samples and to amply VP1 sequence by nested RT-PCR to identify HPeV type.</p><p><b>RESULTS</b>The results showed that 27 of 306 (8.82%) children with acute gastroenteritis were infected HPeV. 11 strains were typed. 9 strains HPeV1, both HPeV2 and HPeV4 was 1 strain. HPeV was mostly identified in autumn season with a peak in July. HPeV seemed relevant in children >2 years old. The range of nucleotide identity between all isolated strains with reference strains was 79%-92%.</p><p><b>CONCLUSION</b>Epidemiology characteristic of HPeV in Jilin was concordance with that of reports. HPeV3 wasnt detected. It's significant to conduct the large scale and long-term surveillance of HPeV.</p>

Pathogens of high incidence of other infectious diarrhea in Guangdong Province, from November 2008 to January 2009 / 病毒学报

From November 2008 to January 2009, a sharp increase of diarrhea in children in Guangdong province appeared, we randomly collected 53 stool specimens from out-patient children with dirrhea in 3 major hospitals (Guangzhou City Children's Hospital, Shenzhen Baoan District Maternal and Child Health Hospital, First Affiliated Hospital of Shantou University). Rotavirus and calicivirus were screened by ELISA and RT-PCR. We found 29 cases of rotavirus infection with diverse serotypes. Only four cases were identified as calicivirus infection. The result indicated that rotavirus was a major pathogen of this high incidence of diarrhea from November 2008 to January 2009 in Guangdong Province.

Analysis on epidemiologic characteristics of viral diarrhea among infants in Taiyuan, Shanxi province, 2007-2008 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the epidemiologic characteristics of virus-induced acute diarrhea in children under 5 years old in Taiyuan, Shanxi province.</p><p><b>METHODS</b>Stool specimens and clinical data were collected from 346 inpatients with acute diarrhea from children less than 5 years old. Rotavirus-positive specimens were identified by ELASA kit. Calicivirus and astrovirus were detected by reverse transcription-polymerase chain reaction (RT-PCR). Adenovirus was done by polymerase chain reaction (PCR).</p><p><b>RESULTS</b>Of the 346 specimens, the percentage of samples with Rotavirus, Calicivirus, Astrovirus, and Adenovirus was 40.8%, 7.5%, 6.4% and 3.2%. Among 141 rotavirus positive samples, serotype G1 (42.6%) was the predominant strain. More than 95% of viral diarrhea patients under hospitalization occurred among children younger than 2 years.</p><p><b>CONCLUSION</b>Rotavirus is the major pathogen contributing to the acute diarrhea. The disease generally peaks at autumn/winter. The predominant rotavirus strain circulated was G1P[8].</p>

Detection of Aichi virus in stool samples from children in Lanzhou / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To identification and analysis Aichi virus from diarrhea and normal children in Lanzhou, and discuss the relationship between Aichi virus and Infant Diarrhea.</p><p><b>METHODS</b>According to the literature published data, Using RT-PCR method to amplified Aichi virus 3CD fragment and the positive products were sequenced and determined, and made the alignment analysis between the nucleotide sequences of the amplified fragment with the known sequence of this virus.</p><p><b>RESULTS</b>There was 1 case detection of Aichi virus in the 46 hospitalized children with diarrhea and 299 children with diarrhea out-patients specifically, Overall detection rate was 0.06%, and there was no Aichi virus was detected in normal control children. 2 viral 3CD gene and the known reference strains of nucleotide sequences were 97%, while phylogenetic analysis showed that genotype of 2 viral belongs to the B.</p><p><b>CONCLUSIONS</b>There existed B Genotype of Aichi virus in China, and more research is needed to clarified the etiology and epidemiology of Aichi virus characteristics.</p>

Sequence the complete sequence of bocavirus I with SISPA-PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).</p><p><b>METHODS</b>To exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I. The virus were enriched through ultracentrifugation. Other nucleic acids, such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.</p><p><b>RESULTS</b>A 4834bp sequence of bocavirus I were assembled.</p><p><b>CONCLUSION</b>SISPA-PCR is an economical and efficient technique for sequence a virus complete genome.</p>

A method of detect the virus which caused diarrhea / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Building a method which can examines virus pathogenic in gastroenteritis excrement specimen.</p><p><b>METHODS</b>Choosing six positive specimens which tested in our laboratory, include adenovirus, calicivirus, rotavirus, bocavirus, astrovirus and enterovirus. Through sequence-independent single primer amplification(SISPA) constructs a gene bank. Looks up the viral gene fragment in gene bank.</p><p><b>RESULTS</b>Obtaining corresponding viral acid sequence in six specimens.</p><p><b>CONCLUSION</b>This research can examine enterovirus and the virus which cause diarrhea, It make a foundation for further studies the viral cause of disease which the examination not yet discovered at present.</p>

Detection of human parechovirus in children hospitalized for acute gastroenteritis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To detect human parechovirus (HPeV) from stool samples of hospitalized children for acute gastroenteritis of undetectable etiology.</p><p><b>METHODS</b>We conducted a real-time PCR to detect HPeV.</p><p><b>RESULT</b>The results showed that 24 of 99 (24%) children with gastroenteritis of undetectable etiology were detected with HPeV. Four known HPeV types (HPeV1, 3, 4, 6) were detected in the present study. HPeV1 (50%) was frequently identified as the predominant strain and follow by HPeV3 (25%), HPeV4 (8.3%) and HPeV6 (4.2%). We were unable to type 3 samples.</p><p><b>CONCLUSION</b>HPeV was prevalent in hospitalized children for acute gastroenteritis of undetectable etiology in China. Further study is needed for clarifying the role of HPeV in gastroenteritis.</p>

Molecular epidemiology of human adenovirus diarrhea among infants and young children in Lanzhou from July 2005 to June 2008 / 中华儿科杂志

<p><b>OBJECTIVE</b>Gastroenteritis is a major cause of childhood morbidity and mortality worldwide. Adenovirus AdV is recognized to be one of the most important pathogens associated with severe dehydrating gastroenteritis. Studies reported elsewhere have shown that about 8%-10% of cases with infantile diarrhea are caused by AdV and in some areas AdV diarrhea even occurred in the form of outbreaks. Studies have confirmed that AdV infections are also very common in infants and young children in China. This study aimed to investigate the molecular epidemiologic characteristics of human adenovirus diarrhea among infants and children with acute diarrhea in Lanzhou, Gansu province, China.</p><p><b>METHOD</b>Stool specimen and case information were collected from both outpatients and inpatients with acute diarrhea in Lanzhou. Polymerase chain reaction was used to detect AdV in stool specimens. The subjects included 709 urban children and 180 rural children, their age ranged from 19 d to 60 months.</p><p><b>RESULT</b>Of the 889 cases, 43 (4.8%) were found positive for AdV. AdV was detected in 14 of 257 (5.4%) cases seen from July 2005 to June 2006, in 4 of 286 cases (1.4%) seen from July 2006 to June 2007. During the period of July 2007 to June 2008, adenovirus was detected in 346 specimens, the positive rate was 7.2% (25/346). AdV detection rates of the three-year period were significantly different. The major AdV subtypes detected were adenovirus (subgenus F) Ad40, Ad41 with a positive rate of 3.8% (34/889), followed by non-enteric adenovirus (Ad12, Ad18, Ad31, Ad2, Ad5, Ad6, Ad7) with a positive rate of 1.0% (9/889) in Lanzhou during the 3 years. Most of the AdV-positive specimens showed Ad41 group F (67.4%, 29/43) as the major epidemic strains, and Ade infection mainly occurred in children under one year of age and no seasonal cluster was found.</p><p><b>CONCLUSION</b>Adenovirus was one of the major etiological agent of viral diarrhea among infants and children in Lanzhou between 2005 and 2008. Ad41 was the prodomiment serotype.</p>

WU polyomavirus and KI polyomavirus detected in specimens from children with acute respiratory tract infection in China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate newly identified polyomavirus WUV and WUV and KIPyV are associated with acute respiratory infections in China, tests were developed to detect WUV and KIPyV gene fragments from nasopharyngeal aspirates collected from children with ARI fron Nov. 2006 to Oct. 2007.</p><p><b>METHODS</b>A total of 318 clinical samples were tested for WUV and KIPyV using PCR method. The positive products were sequenced and compared with those in GenBank.</p><p><b>RESULTS</b>14 of the 318 Samples were positive (WUV was 2.2%, KIPyV was 2.2%). All of children who were positive for WUV or KIPyV had respiratory illness.</p><p><b>CONCLUSION</b>Polyomavirus WU and KIPyV infection may be associated with upper and lower respiratory diseases.</p>

A research on rotaviral diarrhea outbreak in Guangxi Province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze epidemiological characters of an outbreak of rotavirus diarrhea in Daxing County, Guangxi Province.</p><p><b>METHODS</b>Rotavirus-positive specimens were identified by ELISA kit. G/P typing assays were confirmed with multiplex seminested RT-PCR. Full-length VP7 genes of 4 positive specimens were amplified and analyzed.</p><p><b>RESULTS</b>30 cases of Rotavirus-positive were identified from 64 specimens. The attack rate was 46.9%, and G/P typing was G1P[8]. A change of VP7 amino acid residue is at positions 68.</p><p><b>CONCLUSION</b>G1P[8] rotavirus is the etiologic agents of this diarrhea outbreak. In addition, adults were included in this outbreak.</p>