鸡内源性类J亚群禽白血病病毒gp85基因的克隆及分析

为深入探讨J亚群禽白血病病毒(AvianleukosisvirussubgroupJ,ALV J)的亚群特性,利用ALV Jgp85基因两侧的序列片段为引物,从正常SPF蛋鸡、商品肉鸡和DF1细胞基因组中完整地扩增了内源性类ALV Jgp85基因。肉鸡和DF1细胞内源性类ALV Jgp85基因同源性达99 9%;SPF蛋鸡内源性类ALV Jgp85基因与肉鸡和DF1细胞的内源性类ALV Jgp85基因之间同源性达95 6%、95 3%。三种不同来源的内源性类ALV Jgp85基因DNA与IMC10200株ALV J的gp85基因的同源性分别为91 8%、94 1%、94 0%;与ALV J原型株HPRS 103gp85基因的同源性分别为95 6%、98 3%、99 9%。内源性类ALV Jgp85序列与外源性ALV Jgp85基因具有相似或一致的ORF和Jameson Worlf抗原表位优势。

中药联合高效抗反转录病毒疗法对艾滋病患者预后的影响

目的观察中药联合高效抗反转录病毒疗法(highly active antiretroviral therapy,HAART)对艾滋病(acquired immune deficiency syndrome,AIDS)患者预后的影响。方法将2012年10月—2014年10月云南省中医中药研究院防治艾滋病试点项目治疗基地的437例AIDS患者根据临床分期及治疗方法分为ⅡA期空白组(患者拒绝中西药治疗)、ⅡA期中药组(患者拒绝接受HAART治疗,仅采用扶正抗毒丸或康爱保生丸治疗)、ⅡB期西药组(只接受HAART治疗)、ⅡB期中西药结合组(根据辨证接受扶正抗毒丸+HAART或康爱保生丸+HAART治疗)、Ⅲ期西药组及Ⅲ期中西药结合组(治疗方法分别同ⅡB期西药组及ⅡB期中西药结合组)。疗程为24个月。分别在治疗12、24个月时,观察各组AIDS患者发病和死亡人数所占百分比。结果 (1)治疗12、24个月,各组ⅡA、ⅡB期患者所占百分比呈逐渐降低趋势,Ⅲ期(发病期)及死亡患者所占百分比呈逐渐升高趋势,但各组间比较,差异无统计学意义(P>0.05)。(2)ⅡA期患者:与ⅡA期空白组比较,中药组ⅡA期患者所占百分比升高,ⅡB期、Ⅲ期及死亡患者所占百分比降低,差异无统计学意义(P>0.05);(3)ⅡB期患者:与ⅡB期西药组比较,中西药结合组ⅡA、ⅡB期患者所占百分比均升高,但差异亦无统计学意义(P>0.05),Ⅲ期(发病期)死亡患者所占百分比则明显降低(P<0.05);(4)Ⅲ期(发病期)患者:与Ⅲ期西药组比较,中西药结合组ⅡA、ⅡB期患者所占百分比均升高,Ⅲ期(发病期)死亡患者所占百分比降低,差异无统计学意义(P>0.05)。结论无论是否接受治疗、接受何种治疗,AIDS患者发病和死亡人数所占百分比随着时间的推移均呈现逐渐升高的趋势。中西药结合(扶正抗毒丸+HAART疗法或康爱保生丸+HAART疗法)能显著降低ⅡB期死亡患者所占百分比。

Construction of Recombinant Retroviral Vector Containing HIV-1 Tat Gene and Functional Detection of Expressed Tat in Target Cells

Objective: To construct recombinant retroviral vector containing HIV-1 Tatgene and evaluate the junction of the expressed Tat in target cells. Methods: HIV-1 Tat_(101) genewas recovered from pEV plasmid by Hind Ⅲ digestion and cloned into expression plasmid LZESpBMN-Z toconstruct recombinant retroviral expression plasmid named LZRS-Tat_(101). Using the method ofcalcium phosphate, the construct of LZRS-Tat_(101) was then transfected into packaging cell linesPhoenix (ΦNX) which contained env and gal genes encoding structural proteins and pol gene codingfor 3 enzymes ( reverse transcriptase, protease and integrate) essential for retroviral integrationand replication . The stable transfected cell lines was obtained using puromycin to screen for morethan 3 days. Then, immunohistochemical (IHC ) staining was carried out to detect the expressionlevel of Tat_(101) protein in both transiently and stably trancfected ΦNX, respectively. Thesupematants containing recombinant virus collected from transient and stable transfected cells wereemployed to infect 293 cells, respectively, and the expressed Tat in 293 cells was tested by Westernblot. Meantime, the supematants of infected 293 cells was further added to HL3T1 cells which wereHela cell lines containing an HIV-1-LTR/CAT reporter construct to establish a co-culture system.After co-culture for 72 hours, the protein was extracted from HL3T1 cells and used for CAT activityassay. Results: After LZRS- Tat_(101) was transfected into ΦNX, the amount of expressed Tat intransient transfection cells was significantly higher than that in stable transfection cells; Tatcould be detected not only in 293 cells but also in the supematants from 293 cells culture, and Tatin the supematants could activate HIV-1 LTR promoter in HL3T1, resulting in high 'expression of CATlocated at the downstream of LTR. Conclusion: The construct of recombinant retrovirus LZRS-Tat_(101) could express Tat protein in target cells and the expressed Tat was functionally activeand can really exhibit the ability to activate transcription.

一例鹅感染禽白血病的诊断

禽白血病是由禽白血病病毒(ALV) 引起的禽类多种肿瘤性疾病的统称。ALV 为反录病毒科甲型反录病毒属的禽C 型反录病毒群,病毒的天然宿主主要是鸡[1],火鸡、鹌鹑、鸽和鹧鸪也可感染,主要侵害造血系统,导致禽类造血组织中某些细胞大量增生,表现为淋巴细胞性白血病、成红细胞性白血病、成髓细胞性白血病、上皮肿瘤、结缔组织瘤、内皮肿瘤等。鹅感染ALV 的情况也有报道[2]。

和田地区马传贫的防治

马传贫（EIA）由反录病毒科慢病毒亚科中的马传染性贫血病病毒（EIAV）引起的马、骡、驴传染病。其主要发病症状为发烧、贫血、黄疸、出血、心机紊乱,血液学特征性变化为红细胞数、白细胞数、血红蛋白数减少、静脉血中出现吞铁细胞等。马传贫病的特征是病毒的持续感染和临床反复发作,哈尔滨兽医研究所的褚桂芳等的研究表明,病毒的靶细胞是巨噬细胞,病毒在巨噬细胞内增殖,可以避开体液中免疫因子的作用。

异种移植有关的动物传染病

实体器官同种移植是对各种器官末期疾病有效的治疗。在技术再精炼和制定特异有效的免疫抑制方案时,用此方法的受益人数目会继续增长。不幸的是供者来源不能保持,非人类器官的来源成为可能的选择。最近已进行了3例用狒狒和猪的肝脏移植于人的异种移植。很多异种移植的文献着重于技术和免疫问题。对成功移植后传染病的感染危险性却很少彻底地沦述。本文回顾了异种移植可能的传染病危险,并提出研究与预防的可能战略。一些动物的传染病被认为对人可能致命。

Viral kinetics of Enterovirus 71 in human habdomyosarcoma cells

AIM:To characterise the viral kinetics of enterovirus 71 (EV71).METHODS:In this study,human rhabdomyosarcoma (RD) cells were infected with EV71 at different multiplicity of infection (MOI).After infection,the cytopathic effect (CPE) was monitored and recorded using a phase contrast microscope associated with a CCD camera at different time points post viral infection (0,6,12,24 h post infection).Cell growth and viability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both EV71 infected and mock infected cells at each time point.EV71 replication kinet-ics in RD cells was determined by measuring the total intracellular viral RNA with real-time reverse-transcription polymerase chain reaction (qRT-PCR).Also,the intracellular and extracellular virion RNA was isolated and quantified at different time points to analyze the viral package and secretion.The expression of viral protein was determined by analyze the levels of viral structure protein VP1 with Western blotting.RESULTS:EV71 infection induced a significant CPE as early as 6 h post infection (p.i.) in both RD cells infected with high ratio of virus (MOI 10) and low ratio of virus (MOI 1).In EV71 infected cells,the cell growth was inhibited and the number of viable cells was rapidly decreased in the later phase of infection.EV71 virions were uncoated immediately after entry.The intracellular viral RNA began to increase at as early as 3 h p.i.and the exponential increase was found between 3 h to 6 h p.i.in both infected groups.For viral structure protein synthesis,results from western-blot showed that intracellular viral protein VP1 could not be detected until 6 h p.i.in the cells infected at either MOI 1 or MOI 10;and reached the peak at 9 h p.i.in the cells infected with EV71 at both MOI 1 and MOI 10.Simultaneously,the viral package and secretion were also actively processed as the virus underwent rapid replication.The viral package kinetics was comparable for both MOI 1 and MOI 10 infected groups.It was observed that at 3 h p.i,the intracellular virions obviously decreased,thereafter,the intracellular virions began to increase and enter into the exponential phase until 12 h p.i.The total amounts of intracellular virons were decreased from 12 to 24 h p.i.Consistent with this result,the increase of virus secretion occurred during 6 to 12 h p.i.CONCLUSION:The viral kinetics of EV71 were established by analyzing viral replication,package and secretion in RD cells.

Vitamin D improves viral response in hepatitis C genotype 2-3 nave patients

AIM: To examine whether vitamin D improved viral response and predicted treatment outcome in patients with hepatitis C virus (HCV) genotype 2-3. METHODS: Fifty patients with chronic HCV genotype 2-3 were randomized consecutively into two groups: Treatment group [20 subjects, age 48 ± 14 years, body mass index (BMI) 30 ± 6, 65% male], who received 180 μg pegylated α-interferon-2a plus oral ribavirin 800 mg/d (Peg/RBV), together with oral vitamin D3 (Vitamidyne D drops; 2000 IU/d, 10 drops/d, normal serum level > 32 ng/mL) for 24 wk; and control group (30 subjects, age 45 ± 10 years, BMI 26 ± 3, 60% male), who received identical therapy without vitamin D. HCV RNA was assessed by reverse transcription polymerase chain reaction. Undetectable HCV RNA at 4, 12 and 24 wk after treatment was considered as rapid virological response, complete early virological response, and sustained virological response (SVR), respectively. Biomarkers of in? ammation were measured. RESULTS: The treatment group with vitamin D hadhigher BMI (30 ± 6 vs 26 ± 3, P < 0.02), and high viral load (> 400 000 IU/mL, 65% vs 40%, P < 0.01) than controls. Ninety-fi ve percent of treated patients were HCV RNA negative at week 4 and 12. At 24 wk after treatment (SVR), 19/20 (95%) treated patients and 23/30 (77%) controls were HCV RNA negative (P < 0.001). Baseline serum vitamin D levels were lower at baseline (20 ± 8 ng/mL) and increased after 12 wk vitamin D treatment, to a mean level of (34 ± 11 ng/ mL). Logistic regression analysis identifi ed vitamin D supplement [odds ratio (OR) 3.0, 95% CI 2.0-4.9, P < 0.001], serum vitamin D levels (< 15 or > 15 ng/mL, OR 2.2, P < 0.01), and BMI (< 30 or > 30, OR 2.6, P < 0.01) as independent predictors of viral response. Adverse events were mild and typical of Peg/RBV. CONCLUSION: Low vitamin D levels predicts negative treatment outcome, and adding vitamin D to conventional Peg/RBV therapy for patients with HCV genotype 2-3 signifi cantly improves viral response.

Evaluation of a novel hybrid bioartificial liver based on a multi-layer flat-plate bioreactor

AIM: To evaluate the efficacy and safety of a hybrid bioartificial liver (HBAL) system in the treatment of acute liver failure. METHODS: Canine models with acute liver failure were introduced with intravenous administration of D-galactosamine. The animals were divided into: the HBAL treatment group (n = 8), in which the canines received a 3-h treatment of HBAL; the bioartificial liver (BAL) treatment group (n = 8), in which the canines received a 3-h treatment of BAL; the non-bioartificial liver (NBAL) treatment group (n = 8), in which the canines received a 3-h treatment of NBAL; the control group (n = 8), in which the canines received no additional treatment. Biochemical parameters and survival time were determined. Levels of xenoantibodies, RNA of porcine endogenous retrovirus (PERV) and reverse transcriptase (RT) activity in the plasma were detected. RESULTS: Biochemical parameters were significantly decreased in all treatment groups. The TBIL level in the HBAL group was lower than that in other groups (2.19 ± 0.55 mmol/L vs 24.2 ± 6.45 mmol/L, 12.47 ± 3.62 mmol/L, 3.77 ± 1.83 mmol/L, P < 0.05). The prothrombin time (PT) in the BAL and HBAL groups was significantly shorter than the NBAL and control groups (18.47 ± 4.41 s, 15.5 ± 1.56 s vs 28.67 ± 5.71 s, 21.71 ± 3.4 s, P < 0.05), and the PT in the HBAL group was shortest of all the groups. The albumin in the BAL and HBAL groups significantly increased and a significantly higher level was observed in the HBAL group compared with the BAL group (27.7 ± 1.7 g/L vs 25.24 ± 1.93 g/L). In the HBAL group, the ammonia levels significantly decreased from 54.37 ± 6.86 to 37.75 ± 6.09 after treatment (P < 0.05); there were significant difference in ammonia levels between other the groups (P < 0.05). The levels of antibodies were similar before and after treatment. The PERV RNA and the RT activity in the canine plasma were all negative. CONCLUSION: The HBAL showed great efficiency and safety in the treatment of acute liver failure.

Investigation on Infectious Agents of Aborted Pig Fetuses and Its Correlation with PRRSV MLV Vaccine

Infectious agents causing aborted fetus problems in domestic pigs were investigated in this study. More than 10 different infectious agents were known to cause abortion in swine and the major eight viruses among them were inspected. One hundred twelve samples of aborted fetuses from nine provinces in South Korea were collected during April to November, 2013 in this study for the diagnosis of infectious agents causing abortions in pigs. Eight major infection viruses were examined in this study mainly using various diagnostic kits and reverse transcriptase polymerase chain reaction （RT-PCR）. Positive rate of the detection differed from each viruses. In this study, the main focus was the porcine reproductive and respiratory syndrome virus （PRRSV）, which took the second large portion in the positive rate of detection, and then its ORF5 gene was compared with modified live virus （MLV） vaccine strain to figure out the influence of vaccine on disease. Between four positive samples＇ sequence, two of them were 99.9%-100% similar to MLV vaccine strain and two other samples were 88.6%-92.7% similar. Similarity rate of the sequences between the vaccine and virus from aborted fetuses are very crucial, because it implies that abortion in swine can be made due to the usage of vaccine not only by the infection of field virus, and if MLV vaccine actually do have an impact on the infection, usage of the vaccine should be reconsidered.

In vivo Inhibition of NAS Preparation on H9N2 Subtype AIV

NAS preparation, a kind of Chinese herbal medicine found by the Yunnan Eco-agricultural Research Institute, has potential antiviral activity. In this paper, the inhibiting effect of NAS preparation on H9N2 subtype Avian influenza virus (AIV) was investigated in vivo. Chickens infected with H9N2 virus were treated with NAS preparation for 4 days. The virus was then detected by hemoagglutination (HA) test and reverse transcription polymerase chain reaction (RT-PCR). The results showed that no H9N2 virus could be detected at the 7th day when the chickens were treated with 0.2g/kg/d or 0.1g/kg/d of NAS preparation. However the virus could be detected in other chickens without NAS preparation treatment. This result suggested that NAS preparation may be a potential drug candidate to control infection of H9N2 subtype AIV in chickens.

Epigallocatechin gallate inhibits HBV DNA synthesis in a viral replication-inducible cell line

AIM:To analyze the antiviral mechanism of Epigallocatechin gallate(EGCG)against hepatitis B virus(HBV) replication.METHODS:In this research,the HBV-replicating cell line HepG2.117 was used to investigate the antiviral mechanism of EGCG.Cytotoxicity of EGCG was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Hepatitis B virus e antigen(HBeAg)and hepatitis B virus surface antigen(HBsAg)in the supernatant were detected by enzyme-linked immunosorbent assay.Precore mRNA and pregenomic RNA(pgRNA) levels were determined by semi-quantitative reverse transcription polymerase chain reaction(PCR)assay.The effect of EGCG on HBV core promoter activity was measured by dual luciferase reporter assay.HBV covalently closed circular DNA and replicative intermediates of DNA were quantified by real-time PCR assay.RESULTS:When HepG2.117 cells were grown in the presence of EGCG,the expression of HBeAg was suppressed,however,the expression of HBsAg was not affected.HBV precore mRNA level was also downregulated by EGCG,while the transcription of precore mRNA was not impaired.The synthesis of both HBV covalently closed circular DNA and replicative intermediates of DNA were reduced by EGCG treatment to a similar extent,however,HBV pgRNA transcripted from chromosome-integrated HBV genome was not affected by EGCG treatment,indicating that EGCG targets only replicative intermediates of DNA synthesis.CONCLUSION:In HepG2.117 cells,EGCG inhibits HBV replication by impairing HBV replicative intermediates of DNA synthesis and such inhibition results in reduced production of HBV covalently closed circular DNA.

Over Exposure to Efavirenz Plasma Concentrations among Beninese HIV Patients Treated by A 600 mg Daily Dose

Therapeutic plasma concentrations of EFV （efavirenz） are between 1,000 ng/mL and 4,000 ng/mL. Concentrations below 1000 ng/mL are associated with higher risk of treatment failure, and concentrations above 4,000 ng/mL are associated with toxicity. The aim of the study was to appreciate EFV plasmas concentrations profile and the association between plasma levels and various characteristics in Beninese patients treated by a 600 mg standard daily-dose. Blood samples were collected and EFV plasma levels were measured by liquid chromatography coupled with mass spectrometry detection in HIV-infected patients receiving EFV in combination with other antiretroviral drugs for at least 14 days. Adverse effects occurring during treatment were collected through a questionnaire. An over-exposure to EFV among Beninese HIV patients were observed, with 46.4% of patients presenting EFV concentration above 4,000 ng/mL, although adverse effects were tolerated indicating that antiretroviral treatment is safe. The measurement of plasma concentration at the steady-state could contribute to early detection of treatment failure and adapt treatment in subjects presenting serious adverse effects within context of therapeutic drug monitoring.

Detection of the Pandemic H1N1/2009 Influenza A Virus by a Highly Sensitive Quantitative Real-time Reverse-transcription Polymerase Chain Reaction Assay

A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin （HA） genes of the pandemic HlN1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic HIN1/2009 virus infection.

Effect of Maternal Antibodies on the Pathogenesis of Avian Reovirus Infections in Broiler Chickens Using Real-Time Reverse Transcriptase Polymerase Chain Reaction

The effect of maternal antibodies on the pathogenesis of avian reovirus （ARV） was studied in commercial and specific pathogen free broilers （SPF） using a real-time reverse transcriptase （RT）-polymerase chain reaction （PCR） assay, along with the incidence and severity of morbidity, mortality, and gross lesions. ARV RNA was detected in cloacal swabs in both bird groups from the first day throughout the 21 days experiment. Commercial broiler chickens, which had high maternal antibodies against ARV, showed minimum clinical signs, gross lesions, and lower numbers of birds with viral RNA excretion, whereas specific pathogen free （SPF） broiler chickens, which did not have antibody against ARVs, had 30% mortality, more severe signs, and higher numbers of birds excreting viral RNA. The highest peak of SPF birds excreting viral RNA occurred during the time of maximum mortality. The protective effect of maternal antibody on ARV pathogenesis in broiler chickens correlated with the detection of ARV RNA using the real-time RT-PCR.

AA肉种鸡内源性类ALV-J gp85基因及其抗原表位的分析

利用ALV-J gp85基因两侧的序列片段为引物,从山东潍坊某肉种鸡场送检的7日龄肉种鸡基因组中完整地扩增了内源性类ALV-J gp85基因。利用ALV-J env基因特异性引物不能扩增出长度为2.2kb的目的片段,用ALV-J特异性单抗JE-9做间接免疫荧光检测,也呈现阴性反应。这种内源性类ALV-J gp85基因与已报道的禽内源性反录病毒EAV-HP序列的同源性为85.6%～93.8%,其中与ADOL 0系来航鸡EAV-HP的相似性最高(93.8%)。所得到的内源性类ALV-J gp85基因与外源性ALV-J毒株HPRS-103、SD07LK1、NX0101的同源性分别为93.8%、91.8%和91.7%。并且与外源性ALV-J gp85基因具有相似的Jameson-Worlf抗原表位优势。这种内源性类gp85基因很可能与肉雏鸡早期感染ALV-J后所呈现的免疫耐受现象有关。

Factors associated with fatigue in acquired immunodeficiency syndrome patients with antiretroviral drug adverse reactions: a retrospective study

OBJECTIVE： To retrospectively study the preva- lence of fatigue and factors associated with fatigue among acquired immunodeficiency syndrome （AIDS） patients with antiretroviral drug adverse re- actions. METHODS： Data were collected from case reportforms （CRFs） for a project funded by the 1 lth Na- tional 5-year Special Science and Technology Pro- gram on Major Infectious Diseases. Fatigue was de- fined by patient self-report. The outcomes were the prevalence of fatigue and the potential risk factors of fatigue. Univariate and multivariate logistic re- gression analyses were conducted to identify the factors associated with fatigue. RESULTS： Among the 228 subjects, the prevalence of fatigue was 86.8%. In univariate analysis, the sig- nificant differences in demographic characteristics between patients with and without fatigue were： gender [OR=2.29; 95% CI （1.05-4.98）], education lev- el [OR=0.40; 95% CI （0.18-0.85）], anemia [OR=3.80; 95% CI （1.27-11.31）], time of HIV diagnosis [OR= 0.29; 95% CI （0.13-0.65）], and route of infection [OR= 0.14; 9.5% CI （0.06-0.32）]. Abnormal taste and rapid pulse were more commonly seen in patients with fatigue （P〈0.05）, while abdominal distension and lumbar soreness were encountered less often in pa- tients with fatigue （P〈0.05）. Multivariate analysis showed that the four main factors associated with fatigue were anemia [OR=3.S0; 95% CI （1.01 -12.15）], route of infection [OR=3.40; 95% CI （1.21-9.58）; P= 0.02〈0.05], lumbar soreness [OR=0.06; 95% CI （0.02-0.18）; P=0.000〈0.05], and rapid pulse [OR= 10.58; 95% C/（2.16-51.75）; P=0.004〈0.05]. CONCLUSION： This study demonstrated that fa- tigue is common （86.8% prevalence） in AIDS pa- tients with antiretroviral drug adverse reactions, and that anemia, route of infection （i.e., non-com- mercial blood donation） and rapid pulse were risk factors, while lumbar soreness was a protective fac- tor related to fatigue. More attention should bepaid to fatigue and more efforts should be made to find ways to prevent, control and eliminate this symptom in AIDS patients with antiretroviral drug adverse reactions.

Development of a reverse transcription quantitative polymerase chain reaction-based assay for broad coverage detection of African and Asian Zika virus lineages

The Zika virus(ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome(GBS) and microcephaly in humans. The ZIKV is genetically diverse and can be separated into Asian and African lineages. A rapid, sensitive, and specific assay is needed for the detection of ZIKV across various pandemic regions. So far, the available primers and probes do not cover the genetic diversity and geographic distribution of all ZIKV strains. To this end, we have developed a one-step quantitative reverse transcription polymerase chain reaction(qRT-PCR) assay based on conserved sequences in the ZIKV envelope(E) gene. The detection limit of the assay was determined to be five RNA transcript copies and 2.94 × 10^(–3) 50% tissue culture infectious doses(TCID50) of live ZIKV per reaction. The assay was highly specific and able to detect five different ZIKV strains covering the Asian and African lineages without nonspecific amplification, when tested against other flaviviruses. The assay was also successful in testing for ZIKV in clinical samples. Our assay represents an improvement over the current methods available for the detection ZIKV and would be valuable as a diagnostic tool in various pandemic regions.