Rosario Domínguez D, Suárez Morán C M, Rodríguez Roche R, Soler Nodarse M, Guzmán Tirado M G
Instituto de Medicina Tropical Pedro Kourí.
Rev Cubana Med Trop. 1996;48(3):155-60.
4 primer sets, were used to allow the amplification of a nucleotide sequence with unique size for each of the dengue virus serotypes by polymerase chain reaction (PRC). The method consisted in the extraction of ribonucleic acid from supernatant of infected cell cultures, reverse transcription-polymerase chain reaction (RT-PCR). This was completed in approximately 7 hours in a simple tube. The size of the amplified sequence was evidenced by electrophoresis in Agarose gel stained with ethidium. The method showed a sensitivity of at least 2.5 plate forming units (pfu) per tube of reaction. It es useful for the detection and simultaneous identification of the 4 serotypes, starting from supernatants of infected strains cultures from different countries of the Caribbean, Central America, and South America.
使用4组引物,通过聚合酶链反应(PCR)扩增登革病毒各血清型具有独特大小的核苷酸序列。该方法包括从感染细胞培养物的上清液中提取核糖核酸,即逆转录-聚合酶链反应(RT-PCR)。这在一个简单的试管中大约7小时内完成。扩增序列的大小通过用溴化乙锭染色的琼脂糖凝胶电泳来证明。该方法显示每管反应的灵敏度至少为2.5个空斑形成单位(pfu)。它可用于从加勒比地区、中美洲和南美洲不同国家的感染毒株培养物上清液中检测和同时鉴定4种血清型。