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Volume 33, Number 4, December 2000

Using buffy coat for reverse transcriptase-polymerase chain reaction in the diagnosis of dengue virus infection: preliminary study.


Chang-Phone Fung, Yuan-Ming Lee, Benjamin Ing-Tiau Kuo, Su-Pen Yang, Yu-Jiun Chan, Cheng-Yi Liu, Low-Tone Ho, Wu-Tse Liu
Department of Medicine, Taipei Veterans General Hospital and National Yang-Ming University, Taiwan, ROC

 

Methods:

Using reverse transcriptase-polymerase chain reaction (RT-PCR) to detect and type from viremic human serum samples for dengue virus infection is widely used today. However, a few false-negative results were reported due to very low titers of the virus particle in serum samples. As mononuclear cells, macrophages or monocytes are target cells for dengue virus infection, and the replication of virions can be observed in peripheral leukocytes frequently, the amount of virus particle in buffy coat should be higher than those in serum samples. Here, we describe a procedure in which RNA extraction from the buffy coat of a patient with a false-negative serum sample yielded specific viral RNA amplifiable by RT-PCR, thereby providing an alternative choice for the accurate diagnosis of dengue infection.
 



 

J Microbiol Immunol Infect 2000;33:217-222.