OBJECTIVETo develop and optimize the molecular detection assays for recently identified human coronavirus (HCoV) infection.

METHODSBased on the 208 base pair(bp) sequence of novel HCoV reported by HPA of UK, we designed and obtained several pairs of primer (F-1, R-1; F-2, R-2) and Taqman probes (TZ1,TZ2) for detection of novel HCoV. Two of probes were modified with LNA (LNA-TZ1, LNA-TZ2). Then, RT-PCR and various real time RT-PCR assays were developed and optimized in this study. We also compared our assays with the real time RT-PCR assays reported recently by Europe team based on upE or ORF1b target.

RESULTSThe RT-PCR or real time RT-PCR assays for novel HCoV were developed without cross-reactivity with other HCoV and several common respiratory viruses using clinical specimen panel. The analytical sensitivity of assays were less than 50-500 copies per reaction and the detection was improved when Taqman probe modified with LNA-tagged, compared to no LNA-tagged in real time RT-PCR assays. The upE and LNA-TZ1 based assays were better than others.

CONCLUSIONThe molecular detection sensitivity and specificity of TaqMan-based real time PCR assay could be improved when probe tagged with LNA. The upE or LNA-TZ1 based real time RT-PCR assay was recommend for detection of novel HCoV. This study laid a foundation for improving the performance of novel HCoV detection.

Coronavirus ; classification ; genetics ; isolation & purification ; DNA Primers ; genetics ; Humans ; Oligonucleotides ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective: To understand the HIV and syphilis infection and related treatment status of low-fee female sex workers (FSWs) in 3 provinces of China. Methods: Four cross-sectional survey data of low-fee FSWs from six cities (counties) in Guangxi, Yunnan and Hunan Province between October 2012 and July 2015 were obtained from the national science and technology major special project intervention study for reducing sexually transmitted diseases (STDs) and acquired immunodeficiency syndrome (AIDS) in low-fee FSWs' database, which included social demographic characteristics, sexual service characteristics and related medical care seeking behaviors, etc. A total of 2 050 subjects were included in the database. Results: The age of the subjects was (35.16±9.76) years old, with a minimum age of 15 and a maximum age of 67. Those who use condoms every time in commercial sex accounted for 58.9% (n=1 206). Among the reasons of not using condom, the proportion of client reluctant to use was the highest (81.0% (n=682)). Only 38.1% (n=782) was tested for HIV in the last six months. HIV confirmed positive rate was 6.8% (n=139), previous positive accounts for 76.3% (n=106). Rate of antiviral therapy was 55.4% (n=77). By the end of 2015, the loss rate of antivirus treatment was 18.2% (n=14). Those who self-reported symptoms of sexually transmitted diseases in the last 6 months accounted for 9.4% (n=191). 50.3% (n=96) of reporters chose to go to formal hospitals, 23.0% (n=44) chose to go to private clinics and 20.4% (n=39) chose their own medication. The syphilis infection rate was 13.5% (n=277), among them, 91.3% (n=253) were asymptomatic. Conclusion Among low-fee FSWs, the rates of HIV and syphilis infection are higher, the condom consistent use rate, HIV antibodies and syphilis test rate are lower. In this group, active seeking medical idea is poor, the rate of anti-virus treatment and the rate of seeking medical treatment in formal medical institutions is low.

OBJECTIVEFace-to-face interview (FTFI) and polling booth survey (PBS) were applied to compare the high risk behavior among HIV-positive clients of female sex workers (CFSWs).

METHODSThis study was conducted in antiretroviral therapy (ART) out-patients centers in Hengyang city, where clients who had been informed on their HIV-positive status for 6 months or longer were recruited. FTFI was first used to tackle on 8 sensitive questions related to sexual behavior, followed by PBS to poke on the same questions. Results from FTFI and PBS were then compared.

RESULTSCompared with FIFI, results in the PBS showed higher proportion of participants who reported "having had history of sexually transmitted diseases before knowing that they were infected with HIV" (40.6% vs. 24.2%) but lower proportions on "frequency of having sex with regular sexual partner less than 4 times per month" and "using condoms consistently with regular sexual partners in past 6 months (44.4% vs. 63.0%)". There were no significant differences in the remaining 5 questions shown in the results from the two methods.

CONCLUSIONHIV-positive CFSWs continued to practice unsafe sexual behaviors after knowing their HIV related status. Compared with FTFI, PBS seemed to have revealed higher proportion of unsafe sex behaviors, so as to generated more reliable data. While comparing with PBS, the traditional FTFI might have underestimated the risk behaviors among those HIV-positive CFSWs.

Condoms ; utilization ; Data Collection ; methods ; Female ; HIV Seropositivity ; Humans ; Male ; Reproducibility of Results ; Risk-Taking ; Sex Work ; Sex Workers ; Sexual Behavior ; psychology ; statistics & numerical data ; Sexual Partners ; Sexually Transmitted Diseases ; epidemiology ; Unsafe Sex ; psychology

【Objective】 To explore the attention and its relationship with anxiety in patients with glaucoma.【Methods】A prospective case-control study was done on 20 primary open angle glaucoma（POAG）patients and 28 normal subjects. Response time with and without disturbance was examined by using Matlab software. Anxiety was assessed with State Anxiety Inventory（S- AI）and Trait Anxiety Inventory（T- AI）. Response time and inhibiting effects between two groups were compared by t test and Mann- Whitney U test ，respectively. Spearman correlation analysis was applied to determine the relationship between inhibiting effect and anxiety.【Results】The scores of S-AI and T-AI in POAG group were significantly higher than those in normal subjects（t = -3.912，P = 0.001；t = -2.127，P = 0.042）. There were statistically significant differences in the scores between groups even after gender was adjusted by using line regression model（β = 9.449，P = 0.000；β = 7.089，P = 0.018）. The average response time in POAG group（0.84±0.15）s was significantly longer than that in normal participants（0.75±0.14）s（t = -2.127，P = 0.039）. The difference was statistically insignificant when gender was adjusted by using line regression model（β = 0.063，P = 0.210）. The inhibiting effect in POAG group（0.126±0.089）s was longer than that in normal subjects（0.085±0.066）s（Z = -1.715，P = 0.086）. The inhabiting effect wasn′ t statistically significantly correlated with scores of S- AI and T- AI（P＞0.05）.【Conclusions】Patients with POAG showed severer anxiety than normal subjects but attention was comparable. There was no significant correlation between attention and anxiety.