Development of whole-virus multiplex luminex-based serological assays for diagnosis of infections with kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 homologs in macaques.
Development of whole-virus multiplex luminex-based serological assays for diagnosis of infections with kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 homologs in macaques.
复制标题
开发基于全病毒多重 luminex 的血清学测定，用于诊断猕猴中卡波西肉瘤相关疱疹病毒/人疱疹病毒 8 同源物的感染。
DOI：
10.1128/cvi.00673-12
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Rose,TimothyM
中科院分区：
文献类型：
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作者：
Ryan,JonathanT;Rose,TimothyM
卡波西肉瘤相关疱疹病毒（KSHV）/人疱疹病毒8是一种致瘤性鼻状病毒，与所有形式的卡波西肉瘤相关。目前KSHV的血清学检测是基于酶联免疫吸附或免疫荧光测定，其存在各种问题，包括缺乏用于测试比较的定义标准。虽然KSHV是唯一已知的人类鼻状病毒，但在旧世界灵长类动物中发现了两种KSHV样鼻状病毒谱系：RV 1谱系包括猕猴中的KSHV和腹膜后纤维瘤病疱疹病毒（RFHV），RV 2谱系包括来自不同猕猴物种的RRV和MneRV 2。为了建立KSHV相关疾病的动物模型，我们开发了定量多重珠粒血清学检测方法来检测抗鼻状病毒抗原的抗体。将来自KSHV（RV 1）和MneRV 2（RV 2）病毒体的蛋白偶联至光谱上不同的荧光珠，并用于基于Luminex流式细胞术的测定以检测猕猴中的免疫应答。两种检测均显示出大的动态范围，对KSHV和MneRV 2蛋白具有高水平的血清反应性。筛选了来自华盛顿国家灵长类动物研究中心的大量猕猴血清样本，大多数样本（82%）在两种检测中均为阳性，与PCR检测到的高水平RV 1-RV 2共感染一致。猕猴血清对不同的病毒抗原表现出广泛的、可变的和独特的血清学反应，从而可以确定猕猴病毒的初始血清阳性率。Luminex检测试剂盒提供了一种新型的多重方法来评估人类和非人灵长类动物中的鼻病毒感染模式。这将有助于推进我们对鼻病毒生物学和相关宿主免疫反应的理解。
Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 is a tumorigenic rhadinovirus that is associated with all forms of Kaposi's sarcoma. Current serological detection of KSHV is based on enzyme-linked immunosorbent or immunofluorescence assays that suffer from a variety of problems, including the lack of defined standards for test comparison. While KSHV is the only known human rhadinovirus, two lineages of KSHV-like rhadinoviruses are found in Old World primates: the RV1 lineage includes KSHV and retroperitoneal fibromatosis herpesvirus (RFHV) in macaques, and the RV2 lineage includes RRV and MneRV2 from different macaque species. To develop animal models of KSHV-associated diseases, we developed quantitative multiplex bead-based serological assays to detect antibodies against rhadinovirus antigens. Proteins from KSHV (RV1) and MneRV2 (RV2) virions were coupled to spectrally distinct fluorescent beads and used in Luminex flow cytometry-based assays to detect immune responses in macaques. Both assays showed large dynamic ranges with high levels of seroreactivity to both KSHV and MneRV2 proteins. A large set of macaque serum samples from the Washington National Primate Research Center was screened, and most of the samples (82%) were positive in both assays, consistent with the high level of RV1-RV2 coinfection detected by PCR. The macaque sera showed broad, variable, and unique serological responses to the different viral antigens, allowing an initial seroprevalence to be determined for the macaque viruses. The Luminex assays offer a novel multiplexed approach to assess rhadinovirus infection patterns in both humans and nonhuman primates. This will help advance our understanding of rhadinovirus biology and associated host immunological responses.