利用患者外周血白细胞和血清/血浆对登革热感染进行高效诊断。

Efficient diagnosis of dengue infections using patients' peripheral blood leukocytes and serum/plasma.

作者信息

Wang H L, Lin K H, Yueh Y Y, Chow L, Wu Y C, Chen H Y, Sheu M M, Chen W J

机构信息

Department of Clinical Laboratory, Kaohsiung Medical University, Kaohsiung, Taiwan.

出版信息

Intervirology. 2000;43(2):107-11. doi: 10.1159/000025032.

DOI:10.1159/000025032

PMID:10971129

Abstract

OBJECTIVE

Dengue fever has been one of the most important health problems in Taiwan since a large outbreak during 1987 and 1988. It is critically necessary to have a diagnostic approach that can detect early infections in an outbreak or even find infections existing in silent transmission of the disease.

METHODS

To develop an efficient diagnostic protocol, 105 plasma/serum and 35 peripheral blood leukocyte (PBL) specimens from the 1994 outbreak in southern Taiwan were collected for assessment by various diagnostic techniques in this study.

RESULTS

In acute blood samples, dengue viruses were isolated from 19.4% (14/72) and 33.3% (14/42) of reported and confirmed cases, respectively. Viral RNA in serum/plasma was detected from 20.0% (12/60) of acute samples, which was significantly higher than that from convalescent samples (3/44; 6.8%). However, viral RNA in PBLs, detected by reverse transcription polymerase chain reaction (PBL-RT-PCR), could be observed in 73.2% (19/26) and 66.7% (6/9) of acute and convalescent samples, respectively. The persistence of dengue viruses in PBLs was also evidenced by the presence of viral antigens in 42.9% (4/7) of confirmed convalescent samples by the immunofluorescence antibody test. In addition, IgM antibodies were detected in 43.8% (46/105) of reported cases and 85.2% (46/54) of confirmed cases by the IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA).

CONCLUSIONS

Although IgM antibody detection achieved the highest detection rate among techniques assessed in this study, no individual test can actually reach full efficiency for early diagnosis of dengue infections. Here, we propose a protocol which applies MAC-ELISA and PBL-RT-PCR in sequence, by which 22 confirmed cases were definitely proved as dengue positive. High levels of both sensitivity and specificity were shown in this protocol.

摘要

目的

自1987年和1988年台湾地区发生大规模登革热疫情以来，登革热一直是该地区最重要的健康问题之一。迫切需要一种诊断方法，能够在疫情爆发时检测出早期感染，甚至发现疾病隐性传播中的感染情况。

方法

为制定一种有效的诊断方案，本研究收集了1994年台湾南部疫情期间的105份血浆/血清样本和35份外周血白细胞（PBL）样本，采用多种诊断技术进行评估。

结果

在急性血液样本中，报告病例和确诊病例的登革病毒分离率分别为19.4%（14/72）和33.3%（14/42）。血清/血浆中的病毒RNA在20.0%（12/60）的急性样本中被检测到，显著高于恢复期样本（3/44；6.8%）。然而，通过逆转录聚合酶链反应（PBL-RT-PCR）检测，PBL中的病毒RNA在急性样本和恢复期样本中的检出率分别为73.2%（19/26）和66.7%（6/9）。免疫荧光抗体试验在42.9%（4/7）的确诊恢复期样本中检测到病毒抗原，也证明了登革病毒在PBL中的持续存在。此外，通过IgM抗体捕获酶联免疫吸附测定（MAC-ELISA），在报告病例的43.8%（46/105）和确诊病例的85.2%（46/54）中检测到IgM抗体。