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基于安第斯汉坦病毒重组核蛋白的固相酶免疫测定法的开发与评估

Development and evaluation of a solid-phase enzyme immunoassay based on Andes hantavirus recombinant nucleoprotein.

作者信息

Padula P J, Rossi C M, Valle M O Della, Martínez P V, Colavecchia S B, Edelstein A, Miguel S D L, Rabinovich R D, Segura E L

机构信息

Servicio de Biología Molecular, Dpto de Virología, Instituto Nacional de Enfermedades Infecciosas, ANLIS `Dr Carlos G Malbrán', Av. Velez Sarsfield 563, 1281 Buenos Aires, Argentina.

出版信息

J Med Microbiol. 2000 Feb;49(2):149-155. doi: 10.1099/0022-1317-49-2-149.

Abstract

Hantavirus pulmonary syndrome (HPS) with high mortality rate has been reported in five countries in South America. Rapid accurate methods are important both for monitoring acute infections and for epidemiological studies. The Andes virus nucleoprotein amino acid sequence has a high identity percentage compared with other sequences of this region and has been chosen for the development of diagnostic reagents. Andes nucleoprotein expressed in Escherichia coli was applied as antigen in IgG, IgA and mu-capture IgM enzyme-linked inmunosorbent assays (ELISAs). An evaluation of this reagent was conducted to establish its usefulness for differential diagnosis of HPS and seroprevalence studies. Samples from 135 reverse transcription (RT)-PCR-confirmed HPS cases, 77 individuals with other respiratory infections and 957 healthy inhabitants from endemic and non-endemic areas were analysed. The hantavirus-infected patients had an early and strong IgM, IgG and IgA serum antibody response, in most of the cases as early as 1, 7 and 1 days following onset of symptoms, respectively. IgM and IgG detection showed a specificity and sensitivity of 100%. Andes-specific IgM antibodies were found in all patients in the first available sample, which remained detectable for at least 43 days. Specific IgA antibodies were also detected in saliva of patients with acute HPS. The short duration of the disease and the risk for contacts due to person-to-person transmission of Andes virus necessitate the use of highly sensitive tests which might lead to earlier detection of infected people and improve the treatment and management of patients with HPS.

摘要

南美洲五个国家已报告了死亡率很高的汉坦病毒肺综合征(HPS)。快速准确的方法对于监测急性感染和进行流行病学研究都很重要。安第斯病毒核蛋白氨基酸序列与该区域的其他序列相比具有很高的同一性百分比,已被选用于开发诊断试剂。在大肠杆菌中表达的安第斯核蛋白被用作IgG、IgA和μ捕获IgM酶联免疫吸附测定(ELISA)中的抗原。对该试剂进行了评估,以确定其在HPS鉴别诊断和血清流行率研究中的实用性。分析了来自135例经逆转录(RT)-PCR确诊的HPS病例、77例患有其他呼吸道感染的个体以及来自流行和非流行地区的957名健康居民的样本。感染汉坦病毒的患者具有早期且强烈的IgM、IgG和IgA血清抗体反应,在大多数情况下,症状出现后分别最早在第1、7和1天出现。IgM和IgG检测的特异性和敏感性均为100%。在所有患者的首个可用样本中均发现了安第斯病毒特异性IgM抗体,该抗体至少可检测43天。急性HPS患者的唾液中也检测到了特异性IgA抗体。由于安第斯病毒存在人际传播,该疾病病程较短且接触者有感染风险,因此需要使用高度敏感的检测方法,这可能有助于更早地发现感染者,并改善HPS患者的治疗和管理。

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