Kangwanpong D, Bhamarapravati N, Lucia H L
Department of Pathology, Mahidol University, Bangkok, Thailand.
Clin Diagn Virol. 1995 Feb;3(2):165-72. doi: 10.1016/0928-0197(94)00032-p.
The pathogenesis of the severe form of dengue virus infection, dengue hemorrhagic fever, is still obscure. A major research objective has been to determine which body organs are being damaged by dengue virus in this form of dengue. Research has been difficult because dengue hemorrhagic fever is sporadic and tends to occur in parts of the world where modern facilities are scarce and fresh or frozen patient materials are not available. However, major hospitals in these areas have accumulated libraries of paraffin-embedded surgical and autopsy tissues over the years. These tissues may have been subjected to less than optimal fixation and storage. Attempts to localize dengue virus using antigen detection in the stored tissue have encountered many difficulties.
Since viral nucleic acid may be preserved under circumstances which destroy protein antigens, our objective was to detect dengue viral RNA in situ in histologic sections of tissues from patients dying of dengue hemorrhagic fever in Thailand.
Tissues from an 11-year-old boy who died at Ramathibodi Hospital, Bangkok, Thailand in November, 1987 with the clinical diagnosis of dengue hemorrhagic fever were treated by transcribing the dengue viral RNA to DNA followed by amplification using the polymerase chain reaction with subsequent in situ hybridization in order to visualize the cells infected with dengue virus.
Viral RNA was detected in hepatocytes in the mid-zonal region of the liver, as well as scattered macrophages in skin and lymph nodes.
Dengue virus infection can be detected in paraffin-embedded autopsy tissues which have been stored for five years. The same procedure can be used for diagnosing dengue viral infection and for studying the pathogenesis of dengue hemorrhagic fever.
登革病毒感染的严重形式——登革出血热的发病机制仍不清楚。一个主要的研究目标是确定在这种登革热形式下，登革病毒会损害哪些身体器官。研究一直很困难，因为登革出血热是散发性的，并且往往发生在世界上现代设施稀缺、无法获得新鲜或冷冻患者材料的地区。然而，这些地区的主要医院多年来积累了石蜡包埋的手术和尸检组织库。这些组织可能没有得到最佳的固定和保存。在储存的组织中使用抗原检测来定位登革病毒的尝试遇到了许多困难。
由于病毒核酸可能在破坏蛋白质抗原的情况下得以保存，我们的目的是在泰国死于登革出血热患者的组织组织学切片中原位检测登革病毒RNA。
1987年11月在泰国曼谷拉玛蒂博迪医院死亡的一名11岁男孩的组织，临床诊断为登革出血热，通过将登革病毒RNA转录为DNA，然后使用聚合酶链反应进行扩增，随后进行原位杂交，以观察感染登革病毒的细胞。
在肝中区的肝细胞以及皮肤和淋巴结中的散在巨噬细胞中检测到病毒RNA。
在储存了五年的石蜡包埋尸检组织中可以检测到登革病毒感染。相同的程序可用于诊断登革病毒感染和研究登革出血热的发病机制。
