Rabies virus glycoprotein serology ELISA for measurement of neutralizing antibodies in sera of vaccinated human subjects

BackgroundVaccination provides protection against infection by inducing VNAs mainly against RABV surface GP. The measurement of VNAs to RABV is commonly used to assess the level of immunity in humans and animals after vaccination. A VNA titer of ≥ 0.5 IU/mL of sera indicates adequate response to vaccination. Here, we report the development and validation of a RABV GP serology ELISA kit for semi-quantitative measurement of VNA titers in sera of vaccinated human subjects.MethodsUsing a recombinant RABV GP expressed in mammalian cells as the capture antigen, the ELISA method was established using HuMAb NM57 reference initially and HRIG reference subsequently. The limit of detection (LOD), linear range, reproducibility, and precision of the method were examined. Specificity and sensitivity were established to assess the diagnostic accuracy.ResultsRABV GP for ELISA plate coating and optimal dilution of human serum sample was 1 µg/mL and 1:20, respectively. Multiple assays were carried out by different technicians at different laboratories for assay standardization. Using the HRIG reference, the LOD was found to be 0.02–0.06 IU/mL and the linear range was 0.2–10.0 IU/ mL. The inter-assay CVs were in the range of 6.60–10.79%, indicating the reproducibility. None of the 12 known negative human sera, tested positive by ELISA, highlighting the specificity. A total of 415 unknown positive human sera were double-blind tested by the RFFIT and ELISA. The VNA titer cut-off value of ELISA was set at 1.5 IU/mL to ensure no false-positive. The diagnostic specificity and sensitivity were 100% and 91.1%, respectively.ConclusionsThe validation data characterize this ELISA as a suitable method for semi-quantitative measurement of VNA titers in human serum samples to assess vaccination status. The ELISA kit can offer simplicity, speed, low cost and high throughput, making it a practical tool for monitoring the immune response following vaccination.     

Status of vaccine research and development of vaccines for herpes simplex virus

Herpes simplex virus type-1 (HSV-1) and -2 (HSV-2) are highly prevalent global pathogens which commonly cause recurrent oral and genital ulcerations. Less common but more serious complications include meningitis, encephalitis, neonatal infection, and keratitis. HSV-2 infection is a significant driver of the HIV epidemic, increasing the risk of HIV acquisition 3 fold. As current control strategies for genital HSV-2 infection, including antiviral therapy and condom use, are only partially effective, vaccines will be required to reduce infection. Both preventive and therapeutic vaccines for HSV-2 are being pursued and are in various stages of development. We will provide an overview of efforts to develop HSV-2 vaccines, including a discussion of the clinical need for an HSV vaccine, and status of research and development with an emphasis on recent insights from trials of vaccine candidates in clinical testing. In addition, we will touch upon aspects of HSV vaccine development relevant to low and middle income countries.     

Infección por el virus de la hepatitis C y nuevas estrategias de tratamiento

Hepatitis C is a major public health problem worldwide. This disease is caused by the hepatitis C virus, which is characterised by its genetic diversity. The infection is usually asymptomatic. However, between 60% and 80% of HCV-infected individuals will progress to chronic hepatitis, 20% to liver cirrhosis in the medium-to long-term and, each year, between 1% and 4% of these patients with cirrhosis will develop hepatocellular carcinoma (HCC). A Spanish consensus document has recently been drafted to diagnose hepatitis C in a single step, consisting of active investigation (antibodies and viremia) in a single sample, which according to the experts, would reduce the time to access treatment and avoid tracking losses. To definitively change the hepatitis C treatment paradigm, direct-acting antiviral drugs (DAAs) have been approved, whose development has been based on achieving cure rates close to 100% regardless of the genotype of the virus, ie, pangenotypes, with good tolerance and bioavailability. These drugs have constituted a real therapeutic revolution. Supplement information: This article is part of a supplement entitled «SEIMC External Quality Control Programme. Year 2016», which is sponsored by Roche, Vircell Microbiologists, Abbott Molecular and Francisco Soria Melguizo, S.A.© 2019 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosasy Microbiología Clínica. All rights reserved.     

复方倍他米松注射液联合泛昔洛韦胶囊、普瑞巴林胶囊治疗带状疱疹的临床疗效观察

目的 对复方倍他米松注射液联合泛昔洛韦胶囊、普瑞巴林胶囊治疗带状疱疹的临床疗效进行观察并探讨。方法 选取2018 年1 月～2019 年12 月我院收治的118 例带状疱疹患者为研究对象，采用随机抽签的方式将患者分为对照组（泛昔洛韦胶囊+普瑞巴林胶囊+甲钴胺片+维生素B1 片）与观察组（联合复方倍他米松注射液），比较两组治疗总有效率、VAS 评分、QS 评分、C 反应蛋白、无新发皮疹时间、结痂时间及疼痛停止时间、随访2 个月后带状疱疹后遗神经痛发生率以及不良反应发生率。结果 观察组与对照组治疗总有效率分别为94.92%及76.27%，组间对比差异有统计学意义（P＜0.05）；治疗后，观察组VAS 评分显著低于对照组；观察组QS 评分低于对照组（P＜0.05）；治疗后，观察组C 反应蛋白显著低于对照组（P＜0.05）；观察组无新发皮疹时间、结痂时间及疼痛停止时间分别显著短于对照组（P＜0.05）；随访2 个月后，发现观察组带状疱疹后遗神经痛发生率显著低于对照组（P＜0.05）；观察组不良反应发生与对照组比较，差异无统计学意义（P＞0.05）。结论 复方倍他米松注射液联合泛昔洛韦胶囊、普瑞巴林胶囊治疗带状疱疹不仅效果好且安全性高，值得使用。     

Vacunas contra el virus del papiloma humano

Persistent infection by human papilloma virus (HPV) is considered to be the main cause of cervical cancer and other ano-genital cancers. Of more than 30 genotypes able to infect the anogenital tract, it is estimated that, worldwide, HPV 16 and 18 cause 70% of cervical cancers and that HPV 6 and 11 cause more than 90% of genital warts. In the last few years, the morbidity and mortality and health costs associated with cervical cancer and its precursor lesions have stimulated intense research activity to achieve primary prevention of this disease through prophylactic vaccines.     

Infectious meningitis with atypical cerebrospinal fluid cells

Cytologic evaluation of the CSF is often difficult when trying to distinguish between truly neoplastic and reactive cells. Several non-neoplastic conditions may be associated with atypical cells in the CSF, a fact the clinician has to consider to avoid inadequate aggressive theraphies. We report here three patients with infectious meningitis (due to Herpes zoster virus in two, and neuroborreliosis in one) and cytologically atypical cerebrospinal fluid lymphocytes. Further characterization showed that the pleocyrosis in these patients was of reactive origin. Cytomorphology is frequently insufficient and histochemical, immuncytochemical and cellular genome analysis techniques may help differentiate atypical reactive cells from neoplastic cells.     

腺病毒介导的HSV—tk基因治疗大鼠脑胶质瘤实验研究

目的：带有ＨＳＶ－ｔｋ基因的重组腺病毒（ＡｄＨＣＭＶ－ｔｋ）结合核苷类似物（ＮＡ）治疗大鼠Ｃ６脑胶质瘤。方法：用Ｘ－ｇａｌ染色测定ＡｄＨＣＭＶ－ｌａｃＺ转染大鼠Ｃ６胶质瘤细胞的效率。用ＡｄＨＣＭＶ－ｔｋ／ＡＣＶ、ＧＣＶ离体及活体治疗大鼠Ｃ６胶质瘤。结果：ＡｄＨＣＭＶ－ｌａｃＺ感染Ｃ６细胞效率达１００％，ＡｄＨＣＭＶ－ｔｋ感染Ｃ６细胞，在病毒感染复数为１０００时，ＧＣＶ和ＡＣＶ半致死剂量分别为３μｇ／ｍｌ和２０μｇ／ｍｌ，Ａｄ－ＨＣＭＶ－ｔｋ／ＡＣＶ治疗大鼠Ｃ６胶质瘤模型，大鼠生存期超过９０天，而对照组分别为１７．０±１．６天（生理盐水组）、１４．５±１．３天（ＡｄＨＣＭＶ－ｌａｃＺ组），Ｐ＜０．００１。结论：重组腺病毒对靶细胞感染效率可达１００％，ＡｄＨＣＭＶ－ｔｋ用ＧＣＶ的杀伤Ｃ６胶质瘤细胞比ＡＣＶ强，而ＨＳＶ－ｔｋ／ＡＣＶ用腺病毒介导治疗大鼠脑肿瘤疗效显著。     

非鼻咽部T细胞淋巴瘤与EB病毒的相关性

目的：探讨EB病毒（EBV）感染与非鼻咽部T细胞淋巴瘤的关系。方法：用单克隆抗体UCHL-1、L26及EB病毒编码的潜在膜蛋白-1（LMP-1），免疫组化染色确定肿瘤的免疫表型及EB病毒转化蛋白的表达。采用原位杂交方法检测EBV编码的EBERs。结果：21例非鼻咽部T细胞淋巴瘤EBERs5例阳性的（23．8％），其中给内淋巴瘤3例，肺和胃肠淋巴瘤各1例。阳性细胞约占肿瘤细胞的10％～70％。5例EBERs阳性病例中仅1例表达LMP-1，为结内淋巴瘤。结论：非鼻咽部T细胞淋巴瘤可能与EBV的感染有关，LMP-1的阳性率较EBERs低。