The epidemiological characteristics and the source of infection of reemerge human rabies from 2012 to 2017 in Qinghai / 中华疾病控制杂志

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.

Viral and Bacterial Etiology of Acute Febrile Respiratory Syndrome among Patients in Qinghai, China / 生物医学与环境科学（英文）

OBJECTIVE@#This study was conducted to investigate the viral and bacterial etiology and epidemiology of patients with acute febrile respiratory syndrome (AFRS) in Qinghai using a commercial routine multiplex-ligation-nucleic acid amplification test (NAT)-based assay.@*METHODS@#A total of 445 nasopharyngeal swabs specimens from patients with AFRS were analyzed using the RespiFinderSmart22kit (PathoFinder BV, Netherlands) and the LightCycler 480 real-time PCR system.@*RESULTS@#Among the 225 (225/445, 51%) positive specimens, 329 positive pathogens were detected, including 298 (90.58%) viruses and 31 (9%) bacteria. The most commonly detected pathogens were influenza virus (IFV; 37.39%; 123/329), adenovirus (AdV; 17.02%; 56/329), human coronaviruses (HCoVs; 10.94%; 36/329), rhinovirus/enterovirus (RV/EV; 10.03%; 33/329), parainfluenza viruses (PIVs; 8.51%; 28/329), and Mycoplasma pneumoniae (M. pneu; 8.51%; 28/329), respectively. Among the co-infected cases (17.53%; 78/445), IFV/AdV and IFV/M. pneu were the most common co-infections. Most of the respiratory viruses were detected in summer and fall.@*CONCLUSION@#In our study, IFV-A was the most common respiratory pathogen among 22 detected pathogens, followed by AdV, HCoV, RV/EV, PIV, and M. pneu. Bacteria appeared less frequently than viruses, and co-infection was the most common phenomenon among viral pathogens. Pathogens were distributed among different age groups and respiratory viruses were generally active in July, September, and November. Enhanced surveillance and early detection can be useful in the diagnosis, treatment, and prevention of AFRS, as well as for guiding the development of appropriate public health strategies.

Complete genome phylogenetic analysis of five H9N2 avian influenza viruses isolated from poultry flocks in Qinghai lake region / 病毒学报

Five H9N2 avian influenza virus strains were isolated from the environmental samples in live poultry market in Qinghai Lake region from July to September, 2012. To evaluate the phylogenetic characteristics of these H9N2 isolates, the eight gene segments were amplified by RT-PCR and sequenced. The phylogenetic and molecular characteristics of the five strains were analyzed. The results showed that the HA genes of five strains shared 93. 2%-99. 1% nucleotide identities with each other, and the NA genes shared 94. 5%-99. 8% nucleotide identities. The HA cleavage site sequence of the A/environment/qinghai/ 017/2012 isolate was PSKSSRGLF, and the HA cleavage site sequences of the other four strains were all PSRSSRGLF. The HA receptor-binding site had the Q226L mutation. The M1 gene segment had the N30D and T215A mutations. The phylogenetic analysis showed that the five strains were similar to the virus A/chicken/Hunan/5260/2005 (H9N2) isolated in Hunan Province, China and were reassortant genotype viruses; the HA, NA, and NS genes belonged to the Y280-like lineage; the MP gene belonged to the G1-like lineage; the NP, PB1, PB2, and PA genes belonged to the F98-like lineage.

Genetic characteristics of influenza A/H3N2 virus neuraminidase gene: a survey from 2010 to 2012 in Qinghai Province, China / 病毒学报

This study aims to perform a survey of genetic variation in neuraminidase (NA) gene of influenza A/H3N2 virus, as well as related resistance to NA inhibitors, in Qinghai Province of China, 2010 to 2012. Strains of influenza A/H3N2 isolated during an influenza survey from 2010 to 2012 in Qinghai were enrolled by random sampling. Viral RNA was extracted and amplified by RT-PCR. Purified PCR products were sequenced thereafter. Genetic analysis of nucleic acid and the derived amino acid sequences was performed by MEGA 4.0. Phylogenetic trees were also constructed. Strains isolated during 2010-2011 in this study clustered closely with World Health Organization (WHO) 2010-2012 reference vaccine strain A/Perth/16/2009 and 2008-2010 reference vaccine strain A/Brisbane/10/2007 on the phylogenetic tree, while the 2012 isolates were located on another branch. In analysis of derived amino acid sequences, the 2010 isolates mutated at K81T, the 2011 isolates mutated at I26V and D127N, while the 2012 isolates mutated at E41K, P46A, I58V, T71N, L81P, D93G, D127N, D151N, and I307M. The D151N mutation added a glycosylation site to the activity center of NA. No significant variation was discovered in H3N2 NA gene of 2010-2011 isolates in Qinghai, China. Isolates of 2012 were found with significant mutation, which has the potential of inducing minor resistance to NA inhibitors like zanamivir and oseltamivir.

Molecular epidemiological study of measles viruses isolated in Qinghai Province during 2000-2011 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To carry out the molecular epidemiological study of the wild-type measles virus isolated in Qinghai Province during 2000-2011, and provide a scientific basis for the measles elimination.</p><p><b>METHODS</b>Measles viruses were isolated using B95a cell line or Vero/SLAM cell line from throat swabs collected from suspected measles cases during measles outbreak and sporadic in 6 prefectures during 2000-2011. The fragment of 696 nucleotides of N gene carboxy terminal was amplified by using RT-PCR methods. The PCR products were sequenced and analyzed. The phylogenetic tree was conducted with the viruses isolated in viruses from other province.</p><p><b>RESULTS</b>Total 19 measles viruses were isolated during 2000-2011 in Qinghai province and all belong to genotype H1a. The results of phylogenetic tree showed that viruses in 2000-2005 and in 2009-2011 were distributed in two different lineages, and it revealed that these strains belonged to at least 2 viral transmission chains and the viruses circulated during 2000-2005 were not detected after 2005.</p><p><b>CONCLUSION</b>Genotype H1a was the predominant genotype circulated in Qinghai province during 2000-2011. Qinghai measles virus strains had not evolved independently, but coevolved with the measles virus strains in other provinces in mainland China. The variation of important amino acid sites of measles virus should be continuous monitored and provide the scientific strategy for the measles elimination.</p>

Human enterovirus 71 that firstly isolated in Qinghai Province and their genetic features of VP1 region / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genetic characterizations of VP1 gene of human enterovirus 71 (HEV71) isolated from clinical specimens of Hand, Foot and Mouth Disease (HFMD) patients in Qinghai Province in 2008.</p><p><b>METHODS</b>335 clinical samples including stools, throat swabs and vesicle fluids were collected from HFMD patients in Qinghai Province. Viral isolation was performed, and molecular typing was performed with the positive isolates. Then 30 identified HEV71 isolates were performed for entire VP1 coding region amplification and sequencing.</p><p><b>RESULTS</b>Among the 355 clinical samples, 45 human enteroviruses were isolated, and among them, 30 were identified as HEV71. Then 30 HEV71 positive isolates were performed by nucleotide sequencing. It showed that there was some difference in the nucleotide and the amino acid among the 30 HEV71 strains, the homology were 95.2%-100% and 96.6%-100%, respectively. But they all closed to HEV71 strains isolated in China after 1998, and from the phylogenetic tree constructed with 30 Qinghai HEV71 strains and other 35 HEV71 strains represented all known genotype and subgenotype HEV71 strains available from GenBank, it revealed that the 30 Qinghai HEV71 strains clustered within the C4a evolution branch of C4 subgenotype.</p><p><b>CONCLUSION</b>HEV71 was isolated in HFMD patients in Qinghai province, and the HEV71 strains causing HFMD outbreaks in Qinghai province in 2008 were all belong to C4a evolution branch of C4 subgenotype with several transmission chains.</p>