HIV testing uptake and yield among sexual partners of HIV-positive men who have sex with men in Zhejiang Province, China, 2014-2016: A cross-sectional pilot study of a choice-based partner tracing and testing package.

BACKGROUND: Measures to effectively expand tracing and testing to identify undiagnosed HIV infections are significant for the control of HIV/AIDS epidemic among men who have sex with men (MSM). We piloted a choice-based tracing and testing package aimed at improving partner tracing, uptake, and yield of HIV testing for sexual partners of newly diagnosed HIV-positive MSM. METHODS: This package was piloted in the cities of Hangzhou and Ningbo, Zhejiang province, China from June 2014 to June 2016. The package adopted four modes: couples' HIV counseling and testing (CHCT), information assisted partner notification (IAPN), assisted HIV self-testing (HIVST) and patient referral. Data regarding sociodemographic factors and sexual behaviors between HIV-positive MSM and their sexual partners, as well as tracing and testing outcomes of each mode, were collected. RESULTS: Among 2,495 newly diagnosed HIV-positive MSM, 446(18%) were enrolled as index cases (ICs) through two rounds of contact tracing. The ICs disclosed a total of 4,716 sexual partners, of whom 548 (12%) were reachable. The pilot study resulted in a testing uptake of 87% (478/548), and a yield of 16% (74/478) among sexual partners. The generalized linear mixed model showed that the odds of a reachable sexual partner enrolled via IAPN taking an HIV test were 290% greater than that of a partner traced via CHCT (95% CI: 1.6, 9.3). CONCLUSIONS: A choice-based tracing and testing package can feasibly expand HIV testing uptake and case finding among sexual partners of HIV-positive MSM. IAPN may be an acceptable option to reach sexual partners for whom limited contact information is available.

An adult gerbil model for evaluating potential coxsackievirus A16 vaccine candidates.

A suitable animal model of CVA16 infection is crucial in order to understand its pathogenesis and to help develop antiviral vaccines or screen therapeutic drugs. The neonatal mouse model has a short sensitivity period to CA16 infection, which is a major limitation. In this study, we demonstrate that adult (60-day-old) gerbils are susceptible to CVA16 infection at high doses (108.0 TCID50). A clinical isolate strain of CVA16 was inoculated intraperitoneally into adult gerbils, which subsequently developed significant clinical symptoms, including hind limb weakness, paralysis of one or both hind limbs, tremors, and eventual death from neurological disorders. Real-time RT-PCR revealed that viral loads in the spinal cord and brainstem were higher than those in other organs/tissues. Histopathological changes, such as neuronal degeneration, neuronal loss, and neuronophagia, were observed in the spinal cord, brainstem, and heart muscle, along with necrotizing myositis. Gerbils receiving both prime and boost immunizations of alum adjuvant inactivated vaccine exhibited no clinical signs of disease or mortality following challenge by CVA16, whereas 80% of control animals showed obvious clinical signs, including slowness, paralysis of one or both hind limbs, and eventual death, suggesting that the CVA16 vaccine can fully protect gerbils against CVA16 challenge. These results demonstrate that an adult gerbil model provides us with a useful tool for studying the pathogenesis and evaluating antiviral reagents of CVA16 infection. The development of this animal model would also be conducive to screening promising CVA16 vaccine candidates as well as further vaccination evaluation.

Outbreak of HIV Infection Linked to Nosocomial Transmission, China, 2016-2017.

On January 25, 2017, a physician from ZC Hospital in Hangzhou, China, reported to the Zhejiang Provincial Center for Disease Control and Prevention that a potential HIV outbreak might have occurred during lymphocyte immunotherapy (LIT) performed at the hospital on December 30, 2016. We immediately began investigating and identified the index case-patient as an LIT patient's husband who donated lymphocytes for his wife's LIT and later screened HIV-reactive. Subsequent contamination by a technician resulted in the potential exposure of 34 LIT patients. Acute HIV infection was diagnosed in 5 persons. Phylogenetic analysis confirmed that the HIV-1 gag, pol, and env gene sequences from the index and outbreak-related cases had >99.5% similarity. Rapid investigation and implementation of effective control measures successfully controlled the outbreak. This incident provides evidence of a lapse in infection control causing HIV transmission, highlighting the need for stronger measures to protect patients from infectious disease exposure.

Iodine Deficiency in Zhejiang Pregnant Women in the Context of Universal Salt Iodization Programme.

Zhejiang introduced universal salt iodization (USI) programme in 1995 and has achieved the goal of elimination of iodine deficiency disorders (IDD) since 2011. However, no systematical data of iodine nutritional status in population in pregnancy is available. In this cross-sectional study, pregnant women were interviewed to complete questionnaires in addition to handing in samples of urine and household table salt between March 2016 to February 2017. Date of birth, age of pregnancy, ethnicity and dietary iodine habits were recorded. The overall median urinary iodine concentration in 8561 pregnant women was 130.47 µg/L, which was lower than the cut-off value of iodine sufficiency of 150 µg/L recommended by the WHO. Participants using non-iodized salt, taking non-iodine-containing supplements, in coastal and in Han group were independently associated with iodine deficiency. The current USI programme did not supply Zhejiang pregnant women with sufficient iodine intake. They are generally iodine deficient, which have great public health importance since even mild IDD in pregnancy have adverse effects on fetal neurodevelopment. We strongly recommend urgent measures to improve iodine intake in pregnancy.

Reduction in HIV community viral loads following the implementation of a "Treatment as Prevention" strategy over 2 years at a population-level among men who have sex with men in Hangzhou, China.

BACKGROUND: Previous studies have shown that the increased coverage of antiretroviral therapy (ART) could reduce the community viral load (CVL) and reduce the occurrence of new HIV infections. However, the impact on the reduction of HIV transmission among men who have sex with men (MSM) is much less certain. The frequency of HIV infections in MSM have been rapidly increasing in recent years in Hangzhou, China. The "Treatment as Prevention" strategy was implemented at a population-level for HIV-infected MSM from January 2014 to June 2016 in Hangzhou; it aimed to increase the ART coverage, reduce the CVL, and reduce HIV transmission. METHODS: We investigated a subset of MSM diagnosed with HIV pre- and post-implementation of the strategy, using random sampling methods. Viral load (VL) testing was performed for all enrolled individuals; the lower limits of detection were 20 and 50 copies/mL. The data on infections were collected from the national epidemiology database of Hangzhou. Logistic regression analyses were conducted to identify factors associated with the differences in social demographic characteristics and available VL data. RESULTS: The ART coverage increased from 60.7% (839/1383) during the pre-implementation period to 92.3% (2183/2365) during the post-implementation period in Hangzhou. A total of 940 HIV-infected MSM were selected for inclusion in this study: 490 (52.1%) and 450 (47.9%) MSM in the pre- and post-implementation periods, respectively. In total, 89.5% (841/940) of patients had data available on VL rates. The mean CVL was 579 copies/mL pre-implementation and this decreased to 33 copies/mL post-implementation (Kruskal-Wallis < 0.001). The mean CVL decreased for all variables investigated post-implementation of the treatment strategy (P < 0.05). The undetectable VL (≤400 copies/mL) rate pre-implementation period was 50.0% which increased to 84.7% post-implementation (P < 0.001). The mean CVL at the county level significantly decreased in each county post-implementation (Kruskal-Wallis < 0.05). CONCLUSION: Our study confirmed a population-level association between increased ART coverage and decreased mean CVL; overall 84.7% of HIV infected MSM had an undetectable VL and were no longer infectious.

Dietary Patterns and Obesity among Chinese Adults: Results from a Household-Based Cross-Sectional Study.

The key dietary pattern other than dietary factors influencing obesity has been reported by several large epidemiological studies. This study was carried out between 2010 and 2012 including 1613 adult residents in Zhejiang Province. Dietary patterns were extracted by factor analysis based on 24-h dietary recall. Associations with dietary patterns and obesity were examined and adjusted for age and gender by logistic regression. Five dietary patterns were identified by factor analysis with their eigenvalues greater than 1: 'cereal, animal, and plant food', 'high protein food', 'plant food', 'poultry', and 'beverage'. After adjustment for age and gender, the 'cereal, animal, and plant food' and 'beverage' pattern was associated with obesity (OR = 2.924, 3.257; 95% CI = 1.147-7.463, 1.372-7.692). In conclusion, 'cereal, animal, and plant food' and 'beverage' dietary patterns may be associated with increased risk of obesity. 'Cereal, animal, and plant food' dietary patterns may be associated with increased risk of obesity resulting from increased total energy intake by increased protein and fat intake; while a 'beverage' dietary pattern may be associated with increased risk of obesity resulting from increased total energy intake by increased carbohydrate intake. The findings are valuable in targeting future nutrition education.

Prevalence of condomless anal intercourse and recent HIV testing and their associated factors among men who have sex with men in Hangzhou, China: A respondent-driven sampling survey.

Men who have sex with men (MSM) are a large high-risk population for HIV infection in recent years in China. A cross-sectional survey was conducted in Hangzhou, China, to determine rates of condomless anal intercourse (CAI), recent HIV testing (in the recent year) and associated factors using respondent-driven sampling. Questionnaires using face-to-face interviews were employed to collect data on sexual risk behaviors and HIV testing. Five hundred eleven MSM were recruited, of which 459 (89.8%) had anal intercourse in the past 6 months. Of these 459 participants, 457 (99.6%) answered whether they had taken an HIV test in the recent year, so only their data were analyzed. Weighted data were analyzed using bivariate and multivariate logistic regression analysis. The CAI rate with male partners in the past 6 months was 43.7% (95% confidence interval [CI], 34.0-51.5%), while the rate of condomless vaginal intercourse (CVI) was 21.6% (95% CI, 15.6-32.3%). The prevalence of recent HIV testing was 56.8% (95% CI, 48.7-66.5%), while the prevalence of HIV and syphilis were 8.8% and 6.5%, respectively. Multivariate analysis indicated that CAI was associated with earlier homosexual debut, suicidal inclinations, childhood sexual abuse, HIV testing in the recent year, and lower estimate of HIV prevalence. Recent HIV testing was associated with homosexual debut age, engaging in CAI with male partners in the past 6 months, having oral sex in the past 6 months, self-perceived higher likelihood of HIV infection, knowing about antiretroviral therapy for HIV/AIDS, receiving AIDS/sexually transmitted infection (STI) interventions in the past year, and syphilis infection. Given high prevalence of HIV and syphilis, high levels of CAI and CVI, and low HIV testing rate, the results indicated high risk of HIV infection and transmission among MSM. HIV prevention interventions should target MSM with early homosexual debut and psychosocial health problems, while HIV/AIDS education among MSM should focus on increasing knowledge of HIV risk, estimated HIV prevalence and antiretroviral therapy, and improving risk perception of HIV acquisition.

Molecular Epidemiology of Clostridium difficile Infection in Hospitalized Patients in Eastern China.

Few studies on risk factors for and transmission of Clostridium difficile infection (CDI) in China have been reported. A cross-sectional study was conducted for 3 years in eastern China. Consecutive stool specimens from hospitalized patients with diarrhea were cultured for C. difficile. C. difficile isolates from these patients then were analyzed for toxin genes, genotypes, and antimicrobial resistance. A severity score for the CDI in each patient was determined by a blinded review of the medical record, and these scores ranged from 1 to 6. A total of 397 out of 3,953 patients (10.0%) with diarrhea were found to have CDI. Severity of CDI was mild to moderate, and the average (± standard deviation) severity score was 2.61 ± 1.01. C. difficile was isolated from stool specimens in 432 (10.9%) of all the patients who had diarrhea. C. difficile genotypes were determined by multilocus sequence analysis and PCR ribotyping; sequence type 37 (ST37)/ribotype 017 (RT017) (n = 68, 16.5%) was the dominant genotype. Eleven patients (16.2%) with this genotype had a CDI severity score of 5. Overall, three RTs and four STs were predominant; these genotypes were associated with significantly different antimicrobial resistance patterns in comparison to all genotypes (χ2 = 79.56 to 97.76; P < 0.001). Independent risk factors associated with CDI included age greater than 55 years (odds ratio [95% confidence interval], 26.80 [18.76 to 38.29]), previous hospitalization (12.42 [8.85 to 17.43]), previous antimicrobial treatment within 8 weeks (150.56 [73.11 to 310.06]), hospital stay more than 3 days before sampling (2.34 [1.71 to 3.22]), undergoing chemotherapy (3.31 [2.22 to 4.92]), and undergoing abdominal surgery (4.82 [3.54 to 6.55]). CDI is clearly a problem in eastern China and has a prevalence of 10.0% in hospitalized patients. Among risk factors for CDI, the advanced age threshold was younger for Chinese patients than that reported for patients in developed countries.

Prevalence of prior HIV testing and associated factors among MSM in Zhejiang Province, China: a cross-sectional study.

BACKGROUND: Men who have sex with men (MSM) have become one of high-risk population for human immunodeficiency virus (HIV) infection, due to their multiple sex partners and unprotected anal intercourse. Promoting HIV testing is an effective strategy for the prevention and control of HIV infection. We assessed the factors associated with a prior HIV testing history, which could provide guidance for implementation of future HIV intervention programs. METHODS: A cross-sectional study was conducted in three cities of Zhejiang Province, namely, Hangzhou, Ningbo and Wenzhou, using respondent-driven sampling, between December 2013 and June 2014. A face-to-face questionnaire survey was employed to collect relevant information about HIV testing. Univariate and multivariate logistic regression analyses were used to identify the factors associated with a prior HIV testing history. RESULTS: The adjusted rate of prior HIV testing among MSM in Zhejiang Province was 55.9 %. The adjusted rates of HIV and syphilis infections among MSM in Zhejiang Province were 14.0 % and 11.4 %, respectively. A weighted multivariate analysis showed that MSM of older age were more likely to be tested, as were MSM with higher level of education, self-reported homosexuality and a higher frequency of receiving AIDS/sexually transmitted infections educational intervention in the past year. MSM with suicidal inclination and self-perceived lower possibility of HIV infection were less likely to report ever having undergone an HIV test. CONCLUSIONS: The prevalence of prior HIV testing among MSM in Zhejiang Province, China is low. Effective and more frequent interventional measures should be adopted to improve risk awareness and psychosocial support for younger, less educated MSM, and to encourage more MSM to undergo HIV testing.

The Rural-Urban Difference in BMI and Anemia among Children and Adolescents.

There is growing concern over the double burden of over- and under-nutrition in individuals, especially in children and adolescents, which could dwarf their growth and development. This study aims to explore the rural-urban difference in BMI and anemia among children and adolescents. A stratified cluster sampling technique was employed. Dietary data were collected through interviews, and anthropometric values were measured. There were 1534 children and adolescents who participated in this study, including 775 male and 759 female participants. The prevalence of obesity among children living in a city, township and rural area was 10.3%, 8.5% and 5.5%, and that among adolescents was 1.4%, 2.9% and 2.8%. The prevalence of anemia among children and living in a city, township and rural area was 4.3%, 2.5% and 4.5%, while that among adolescents was 6.1%, 3.7% and 11.3%, respectively, with significant difference (χ² = 10.824, p = 0.004). The prevalence of being overweight, obesity and anemia was significant when comparing children with adolescents (χ² = 37.861, p = 0.000; χ² = 19.832, p = 0.000; χ² = 8.611, p = 0.003). Findings of this study indicate the double burden of malnutrition in Zhejiang province, characterized by a high prevalence of being overweight, obesity and anemia among children and a high prevalence of anemia among adolescents living in townships.

US Gulf-like toxigenic O1 Vibrio cholerae causing sporadic cholera outbreaks in China.

OBJECTIVES: Cholera is potentially a life threatening disease caused by toxigenic Vibrio cholerae. Here we report the identification and characterisation of 76 non-7th pandemic clone O1 V. cholerae isolates including 65 clinical isolates from diarrhoeal patients from 2005 to 2014 in Zhejiang Province, China. METHODS: We used multilocus sequence typing (MLST) to characterise 65 V. cholerae isolates. Pulse-Field Gel Electrophoresis (PFGE) was performed on a subset of the isolates and whole-genome sequencing was done on 13 isolates. RESULTS: MLST separated 65 isolates into 19 sequence types (STs). Thirty three isolates belonged to ST75 which also contains the US Gulf Coast clone. PFGE separated the 33 isolates into 16 pulsotypes. Whole genome sequencing of 10 ST75 isolates showed that the US Gulf Coast clone and the Chinese ST75 isolates can be separated into two distinct lineages, ST75a and ST75b. All Zhejiang ST75 isolates were ST75b. CONCLUSION: PFGE and genome sequencing confirmed the linked cases and identified small outbreaks caused by ST75b. The emergence and potential spread of ST75b may pose significant threat to public health. Epidemiological surveillance is required to further understand its epidemic potential.

Risky sexual networks and concentrated HIV epidemics among men who have sex with men in Wenzhou, China: a respondent-driven sampling study.

BACKGROUND: The high and continually increasing prevalence of human immunodeficiency virus (HIV) and other sexually transmitted diseases among men who have sex with men (MSM) in China underscores the critical importance of examining the exact sexual networks that result in HIV transmission, as well as HIV infection, using powerful sampling methods, such as respondent-driven sampling (RDS), to improve the sexual health of this population. METHODS: Using RDS, a cross-sectional study was conducted among MSM in Wenzhou, Zhejiang province, China from December 2013 to June 2014. The type of sex, numbers of anal sex partners, male oral sex partners and vaginal sex partners, condom use during each type of sex over the previous 6 months, prevention behaviors, risk perception, and the burdens of HIV and syphilis were investigated and analyzed. RESULTS: Of 424 MSM, a great number of them did anal sex, male oral sex, and vaginal sex during the previous 6 months, and weighted estimates for the prevalence that MSM did not conduct these sexual behaviors were 11.2 % (95 % confidence interval [CI] =6.7-16.50 %), 20.3 % (95 % CI = 15.2-27.1 %), and 58.9 % (95 % CI = 52.1-65.8 %), respectively. Multiple sexual partners, engaging in regular, casual and commercial sex, and lack of condom use during all types of sex were common among MSM. The estimated HIV and syphilis prevalences were 22.8 % (95 % CI = 16.9-28.5 %) and 9.7 % (95 % CI = 6.4-13.6 %), respectively. Of the participants, 53.5 % (95 % CI = 45.3-60.2 %) received HIV-related interventions during the previous year, 48.1 % (95 % CI = 39.7-55.1 %) had never been tested for HIV, and only 14.1 % (95 % CI =10.1-19.2 %) perceived a risk of contracting HIV. Multiple logistic regression analysis revealed that age over 44 years (adjusted odds ratio [AOR] = 3.60, 95 % CI = 1.34-9.64), a monthly income of 3001-4000 yuan (approximately 470-630 US$) (AOR = 1.96, 95 % CI = 1.67-3.60), multiple anal sex partners (AOR = 1.93, 95 % CI = 1.15-3.24), awareness of the possibility of contracting HIV (AOR = 3.18, 95 % CI = 1.56-6.48), and current syphilis infection (AOR = 3.01, 95 % CI = 1.44-6.29) were predictors of HIV infection. CONCLUSIONS: HIV transmission has become highly prevalent and will likely become more prevalent among MSM and their female partners if these risky sexual networks persist. Our findings call for urgent and effective interventions to prevent the rapid transmission of HIV among MSM in Wenzhou.

High prevalence of HIV among men who have sex with men in Zhejiang, China: a respondent-driven sampling survey.

OBJECTIVES: To examine the prevalence of HIV and risk behaviours among men who have sex with men (MSM) and to explore the feasibility of using respondent-driven sampling in this population in order to conduct customised interventions among MSM in the future. DESIGN: Cross-sectional study. SETTING: Zhejiang, China. PARTICIPANTS: 1316 MSM. PRIMARY AND SECONDARY OUTCOME MEASURES: HIV prevalence rates and factors associated with HIV infection; sociodemographic and behavioural information of participants, syphilis prevalence rates. RESULTS: The adjusted prevalence of HIV and syphilis were 13.8% and 11.4%, respectively. Multivariate analysis showed that higher educational level, support treatment of HIV, negative syphilis are protective factors of HIV infection. MSM who had heterosexual behaviour before and whose primary sexual partner was HIV-positive were less likely to be infected with HIV compared with their counterparts, while frequency of sexual behaviour with primary sexual partner was positively associated with HIV infection. CONCLUSIONS: This survey confirmed a high HIV prevalence among MSM in Zhejiang province. MSM are extremely vulnerable to HIV infection and comprehensive interventions are urgently needed to slow the spread of HIV among MSM.

Rapid HIV antibody testing among men who have sex with men who visited a gay bathhouse in Hangzhou, China: a cross-sectional study.

OBJECTIVE: To understand the prevalence and correlates of rapid HIV antibody testing (RHT) among men who have sex with men (MSM) clients of gay bathhouses. DESIGN: Cross-sectional questionnaire survey. SETTING: This study was conducted in a gay bathhouse in Hangzhou, China. PARTICIPANTS: 354 MSM were validly recruited from October to December 2012. Inclusion criteria were (1) men who visited the gay bathhouse, (2) men who had engaged in sex with men during the previous 6 months, (3) first-time participants in this survey and (4) men who were HIV-negative if already tested. MEASURES: Sociodemographic measures included factors related to sexual behaviour and HIV risk perception, and the scales of HIV-related knowledge and behavioural intervention that each participant received. RESULTS: Of the 354 participants, 222 (62.7%) were rapid tested during the previous 6 months; of them, 66.2% were tested at the Centers for Disease Prevention and Control (CDC), and 46.8% at gay venues. The following factors were independently associated with rapid testing within the previous 6 months: sexual initiation at 20-29 years of age, ever having undergone standard testing, ever having seen a sexually transmitted disease doctor, consistent use of condom during the past 6 months, familiarity with RHT and perception of possible HIV infection. CONCLUSIONS: Publicity of RHT and risk education for HIV infection are necessary to promote RHT among MSM who visit gay bathhouses. The characteristics of sexual behaviours among those who do and do not undergo RHT should be taken into consideration while promoting the service in this group.

Genetic Susceptibility Is One of the Determinants for Severe Fever with Thrombocytopenia Syndrome Virus Infection and Fatal Outcome: An Epidemiological Investigation.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease in China and case-fatality rate of SFTS is very high (approximately 10%). However, genetic susceptibility for SFTS virus (SFTSV) infection and fatal outcome of SFTSV infection in humans are unclear. In this study, we investigated the clinical, laboratory and epidemiological features of SFTS in a cluster of three sisters who died of SFTSV infection between late April and mid-May 2014. Before disease onset, two of the sisters (Case A and case B) had common exposure history for ticks by working together in a field to pick tea leaves from April 8 to April 12. The third sister (Case C) did not live or work together with case A and B, but had ticks in her living environment. SFTSV RNA sequences were amplified from three cases were not identical, suggesting that the three sisters were most likely infected with SFTSV through tick bite rather than through person-to-person transmission of SFTSV. The sequence of SFTSV from case C was identical to SFTSV sequences from 3 groups of ticks collected around the residential area of case C. Seroprevalence of SFTSV IgG antibody among healthy population in the area where the patients resided was 4.05% (3/74). The majority of SFTSV infections were mild cases and all three sisters died of SFTSV infection suggested that they were highly susceptible to SFTSV. Our findings indicated that genetic susceptibility was a risk factor for SFTSV infection and fatal outcome.

Enterovirus 71 infection causes severe pulmonary lesions in gerbils, meriones unguiculatus, which can be prevented by passive immunization with specific antisera.

Neurogenic pulmonary edema caused by severe brainstem encephalitis is the leading cause of death in young children infected by Enterovirus 71 (EV71). However, no pulmonary lesions have been found in EV71-infected transgenic or non-transgenic mouse models. Development of a suitable animal model is important for studying EV71 pathogenesis and assessing effect of therapeutic approaches. We had found neurological disorders in EV71-induced young gerbils previously. Here, we report severe pulmonary lesions characterized with pulmonary congestion and hemorrhage in a gerbil model for EV71 infection. In the EV71-infected gerbils, six 21-day-old or younger gerbils presented with a sudden onset of symptoms and rapid illness progression after inoculation with 1×105.5 TCID50 of EV71 via intraperitoneal (IP) or intramuscular (IM) route. Respiratory symptoms were observed along with interstitial pneumonia, pulmonary congestion and extensive lung hemorrhage could be detected in the lung tissues by histopathological examination. EV71 viral titer was found to be peak at late stages of infection. EV71-induced pulmonary lesions, together with severe neurological disorders were also observed in gerbils, accurately mimicking the disease process in EV71-infected patients. Passive transfer with immune sera from EV71 infected adult gerbils with a neutralizing antibody (GMT=89) prevented severe pulmonary lesion formation after lethal EV71 challenge. These results establish this gerbil model as a useful platform for studying the pathogenesis of EV71-induced pulmonary lesions, immunotherapy and antiviral drugs.

Isolation and characterization of H7N9 avian influenza A virus from humans with respiratory diseases in Zhejiang, China.

In 2013, the novel reassortant avian-origin influenza A (H7N9) virus was reported in China. Through enhanced surveillance, infection by the H7N9 virus in humans was first identified in Zhejiang Province. Real-time reverse-transcriptase-polymerase-chain-reaction (RT-PCR) was used to confirm the infection. Embryonated chicken eggs were used for virus isolation from pharyngeal swabs taken from infected human patients. The H7N9 isolates were first identified by the hemagglutination test and electron microscopy, then used for whole genome sequencing. Bioinformatics software was used to construct the phylogenetic tree and for computing the mean rate of evolution of the HA gene in H7Nx and NA in HxN9. Two novel H7N9 avian influenza A viruses (A/Zhejiang/1/2013 and A/Zhejiang/2/2013) were isolated from the positive infection cases. Substitutions were found in both Zhejiang isolates and were identified as human-type viruses. All phylogenetic results indicated that the novel reassortant in H7N9 originated in viruses that infected birds. The sequencing and phylogenetic analysis of the whole genome revealed the mean rate of evolution of the HA gene in H7NX to be 5.74E-3 (95% Highest posterior density: 3.8218E-3 to 7.7873E-3) while the NA gene showed 2.243E-3 (4.378E-4 to 3.79E-3) substitutions per nucleotide site per year. The novel reassortant H7N9 virus was confirmed by molecular methods to have originated in poultry, with the mutations occurring during the spread of the H7N9 virus infection. Live poultry markets played an important role in whole H7N9 circulation.

Development of novel AllGlo-probe-based one-step multiplex qRT-PCR assay for rapid identification of avian influenza virus H7N9.

Recently, human deaths have resulted from infection with low-pathogenicity avian influenza virus H7N9 strains that have emerged recently in China. To strengthen H7N9 surveillance and outbreak control, rapid and reliable diagnostic methods are needed. To develop a sensitive quantitative real-time RT-PCR assay for rapid detection of H7N9 viral RNA, primers and AllGlo probes were designed to target the HA and NA genes of H7N9. Conserved sequences in the HA and NA genes were identified by phylogenic analysis and used as targets for H7N9 virus detection. The similarities of the targeted HA and NA gene sequences from different H7 and N9 influenza virus strains were 93.2-99.9 % and 96.0-99.6 %, respectively The specificity and sensitivity of the new multiplex real-time qRT-PCR was established. The test was used for the detection of viral RNA in human pharyngeal swabs and environmental samples. The detection limit of the multiplex qRT-PCR was estimated to be about 10(-1) TCID50/reaction. Finally, the diagnostic sensitivities of the multiplex qRT-PCR, virus isolation and TaqMan qRT-PCR were compared using pharyngeal swabs and environmental samples. These analyses yielded positive results in 46.7 %, 43.3 % and 20.0 % of the samples, respectively. The novel multiplex AllGlo qRT-PCR is a rapid and sensitive method to identify H7N9 virus in clinical and environmental samples and can be used to facilitate studies on the epidemiology of H7N9 virus.

The effects of different dosage levels of hepatitis B vaccine as booster on anti-HBs-negative children 5-15 y after primary immunization; China, 2009-2010.

The changes in lgG antibody levels to hepatitis B surface antigen (HBsAg) and in antibody to HBsAg (anti-HBs) seroconversion rates due to different dosages of hepatitis B vaccine (HepB) were compared in 2106 children. Children who had been previously vaccinated as infants with HepB were revaccinated at 5-15 y of age, after which the antibody titers were determined. After the first booster dose, the anti-HBs seroconversion rate (defined as an anti-HBs ≥10 mIU/ml) with 10 µg of HepB (93.6%) was significantly greater than the rate with 5 µg of HepB (90.3%) (P<0.05); the anti-HBs seroconversion rate in 10-15-y-old boys vaccinated with 10 µg of HepB (90.9%) was significantly greater than the rate with 5 µg of HepB (84.3%) (P<0.05). The anti-HBs seroconversion rates after the third booster dose with 5 or 10 µg of HepB were greater than those after the first booster dose (99.6% and 99.7%, vs. 90.3% and 93.6%, P<0.05); as was the corresponding GMTs (658 ± 4 mIU/ml and 2599 ± 3 mIU/ml, vs. 255 ± 11 mIU/ml and 877 ± 11 mIU/ml [P<0.05]). The immunization effects of booster vaccination with 3 doses of HepB with 5 or 10 µg are effective; a single booster dose with 10 µg of HepB for 10-15-y-old boys and with 5 or 10 µg of HepB for 5-9 y old boys and for 5-15-y-old girls are effective in generating protective antibody against HBV; however, for anti-HBs-negative children after a single dose of booster, 3 doses are needed.