病毒血清学检测与核酸检测在输血传染病筛查中的应用价值

目的:探讨病毒血清学检测与核酸检测在输血传染病筛查中的应用价值。方法:选取2016年2月至2019年5月于我院无偿献血的4000名志愿者的血样样本,所有血液样本均接受分离血清,对血清进行病毒血清学检测及核酸检测,以血培养结果为金标准。对比病毒血清学检测、核酸检测单独检测与联合检测的阳性检出率及符合率。结果:联合检测与血培养阳性率差异无统计学意义(P>0.05);联合检测阳性率高于病毒血清学检测、核酸检测,差异具有统计学意义(P<0.05);联合诊断准确率高于病毒血清检测准确率、核酸检测准确率,差异具有统计学意义(P<0.05)。结论:病毒血清学检测与核酸检测在输血传染病筛查中具有较高使用价值,两者联合诊断准确性更高,可以作为临床输血传染病筛查的有效手段,值得推广。

病毒血清学检测与核酸检测在输血传染病筛检中的应用

目的探讨病毒血清学检测与核酸检测在输血传染病筛检中的应用价值。方法选取2016—2018年的1 600名无偿献血人员作为研究对象,均应用病毒血清学检测和核酸检测筛检,观察检测结果。结果病毒血清学检测出乙型肝炎1例,艾滋病1例,阳性检出率为0. 125%;核酸检测出丙型肝炎1例,艾滋病1例,阳性检出率为0. 125%;两种技术联合筛检,乙型肝炎1例,丙型肝炎1例,艾滋病1例,阳性检出率为0. 188%。两种技术联合筛检阳性检出率显著高于单独病毒血清学检测和核酸检测,差异有统计学意义(P <0. 05)。结论病毒血清学检测与核酸检测在输血传染病筛检中均具有一定的筛检价值,两者联合应用能够提高阳性检出率。

病毒血清学检测和优生健康教育对母婴的影响

目的探析病毒血清学检测和优生健康教育对母婴的影响。方法回顾性分析2017年10月至2018年10月于宁陵县妇幼保健院就诊的431例孕妇的临床资料,根据其产前是否接受优生健康教育分为对照组(210例)和观察组(221例)。所有孕妇均接受病毒相关免疫标志物检测,观察组孕妇夫妇接受3个月的优生健康教育。观察两组孕妇病毒相关免疫检测结果,比较两组孕妇优生健康知识水平和新生儿出生缺陷率。结果431例孕妇中,病毒血清学检测总阳性率为17.9%(77/431),其中乙型肝炎病毒(HBV)阳性率最高,为13.2%(57/431)。初次就诊时,对照组和观察组孕妇健康知识调查问卷得分分别为(46.5±7.5)、(47.1±8.7)分,差异无统计学意义(P>0.05)。预产期前,对照组和观察组孕妇健康知识调查问卷得分分别为(65.2±9.7)、(86.5±11.4)分,观察组得分高于对照组,差异有统计学意义(P<0.05)。对照组和观察组出生缺陷率分别为3.33%(7/210)、0.45%(1/221),观察组新生儿出生缺陷率低于对照组,差异有统计学意义(P<0.05)。结论在进行病毒血清学检测的同时,对孕妇夫妇进行优生优育健康教育,有助于提高孕妇健康知识水平,减少新生儿出生缺陷的发生。

病毒血清学检测及核酸检测技术用于输血传染病筛检中的价值评价

目的:研究病毒血清学检测以及核酸检测技术用于输血传染病筛检中的临床价值。方法:选取在2018年7月至2018年11月进行无偿献血的1400名社会人群作为本次研究对象,参与研究的人群均采用病毒血清学以及核酸检测技术,并对检测结果进行观察以及对比。结果:结果显示,病毒血清学检测出乙型肝炎、艾滋病都为1例,核酸技术检测出乙型肝炎、艾滋病各1例,当两种技术合用之后检测出乙型肝炎、丙型肝炎以及艾滋病各1例。两种技术合用之后阳性检测率明显高于单独使用病毒血清以及核酸检测方法。结论:病毒血清学检测与核酸检测在输血传染病筛检中都具有一定的临床价值,但是两者合用之后能够提高传染病的阳性检测率。

A Virus-type Specific Serological Diagnosis of Flavivirus Infection Using Virus-like Particles

Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus （WNV）, dengue virus （DENV）, yellow fever virus （YFV）, and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test （PRNT） is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety levet-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles （VLPs） that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps： （i） VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37℃ for 1 h; （ii）the neutralized VLPs are used to infect Vero cells; and （iii） the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen （as quantified by the luciferase activities） is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the ＂gold standard＂ of the classic PRNT; importantly, it shortens the assay time from 〉10 days to 〈1 day, and can be performed in biosafety level-2 facility.

Simultaneous Detection of Three Arboviruses Using a Triplex RT-PCR Enzyme Hybridization Assay

Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation. We developed a cost-effective, rapid, and highly sensitive one-step "triplex RT-PCR enzyme hybridization" assay for simultaneous detections of Japanese Encephallitis virus (JEV, Flaviviridae), Getah virus (GETV, Togaviridae), and Tahyna virus (TAHV, Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction. The analytical sensitivity of this assay was 1 PFU/mL for JEV, 10 PFU/mL for GETV, and 10 PFU/mL for TAHV. This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods. When "triplex RT-PCR enzyme hybridization" was applied to 29 cerebrospinal fluid (CSF) samples that were JEV-positive by normal RT-PCR assay, all samples were strongly positive for JEV, but negative for GETV and TAHV, demonstrating a good sensitivity, specificity, and performance at CSF specimen detection.

Serological Detection of Tomato Pepino mosaic virus in Morocco

Pepino mosaic virus （PepMV）, monopartite RNA virus, 6,500 pb, belonging to Flexiviridae and Potexvirus group, is highly infectious and easily transmissible. Its economic impact is major for the tomato producer＇s countries. Prevention, based on early virus detection is the only effective control measure. Monoclonal antibodies appeared to be very useful tool. The authors used for the production of monoclonal antibodies hybridomas technique, by fusing spleen cells of immunized BALB/c mice to PepMV and SP2/O cancerous cells. The aim of this work is to produce hybridomas producers of Mab that could be used for ELISA in Morocco. At the same time, these efforts will serve to decrease expenses of producers concerning phytosanitory control. We obtained 16 hybridomas lines producers of Mab specific for PepMV. They were tested for efficiencies in ELISA and two lines were retained for production of Mab on large scale （1B 11-G 10 and 5A l-G5）. Isotyping of these two lines showed that they are belonging to IgG 1 class and easily purified by affinity chromatography in agarose column by protein A. The conjugation of these two antibodies to alkaline phosphatase has been verified by DAS-ELISA. These antibodies will enable to diagnose the disease from infected tomato plants, integrating several serological tests to control it and target the actions of struggles.

Monoclonal Antibody-Based Serological Detection Methods for Wheat Dwarf Virus

Wheat dwarf disease caused by wheat dwarf virus(WDV) is currently present in wheat growing regions in China and causes serious losses in wheat yield. To develop reliable and effective serological detection methods for WDV, the coat protein(CP) gene of WDV was cloned and expressed in Escherichia coli. The purified recombinant CP protein was immunized to BALB/c mice, and four hybridoma cell lines(i.e. 18G10, 9G4, 23F4 and 22A10) secreting anti-WDV monoclonal antibodies(MAbs) were obtained through the hybridoma technique. Using the prepared MAbs, an antigencoated-plate enzyme-linked immunosorbent assay(ACP-ELISA) and a dot-ELISA were established for detecting WDV in wheat samples. The most sensitive ACP-ELISA based on MAb 23F4 or 22A10 was able to detect WDV in 1:163,840(w/v,g/mL) diluted WDV-infected wheat plant crude extracts. The dot-ELISA based on MAb 23F4 was the most sensitive and able to detect the virus in 1:5,120(w/v, g/mL) diluted wheat plant crude extracts. A total of 128 wheat samples were collected from wheat growing regions in the Shaanxi and Qinghai provinces, China, and were screened for the presence of WDV using two developed serological assays. Results from the survey showed that approximately 62% of the samples were infected with WDV. PCR followed by DNA sequencing and sequence alignment validated the results from the two serological assays. Therefore, we consider that these two serological detection methods can be significantly useful for the control of WDV in China.