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为了早期快速诊断黄病毒感染，在其ＮＳ１基因序列设计了一对通用引物，扩增序列长度为４１３ｂｐ；在登革病毒（ＤＥＮ）１，２，３，４型和乙型脑炎病毒（ＪＥＶ）设计了各型的内引物，扩增序列长度分别是ＤＥＮ１型为２６２ｂｐ，ＤＥＮ２型为１８９ｂｐ，ＤＥＮ３型为３９２ｂｐ，ＤＥＮ４型为９７ｂｐ，ＪＥＶ为３２３ｂｐ。应用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）成功地扩增了ＤＥＮ１－４型和ＪＥＶ基因部分片段，其扩增片段的大小与设计相符合。应用套式ＰＣＲ检测临床诊断为登革热的患者血清标本７８份，证实ＤＥＮ１型阳性１８份，ＤＥＮ２型阳性４８份，其中８份同时合并感染ＤＥＮ４型。采用套式ＰＣＲ检测临床诊断为乙型脑炎患者血标本４２份，证实乙型脑炎病毒感染者３５份。结果表明，该法能直接检测发病患者早期血标本中的病毒基因，并在２天内完成对黄病毒的鉴别诊断。
In order to rapidly diagnose flavivirus infection in early stage, a pair of universal primers was designed based on its NS1 gene sequence. The amplified sequence length was 413 bp. In the dengue virus (DEN) 1, 2, 3, 4 and JEV ) Designed various types of primers, the length of the amplified sequences were DEN1 type was 262bp, DEN2 type was 189bp, DEN3 type was 392bp, DEN4 type was 97bp, JEV was 323bp. The cDNA fragments of DEN1-4 gene and JEV gene were successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR). The size of amplified fragment was in accordance with the design. 78 samples of dengue fever were detected by nested PCR, 18 of which were positive for DEN1 and 48 were positive for DEN2, of which 8 were infected with DEN4 at the same time. Using nested PCR detection of 42 cases of blood samples of patients with Japanese encephalitis, 35 cases of Japanese encephalitis virus were confirmed. The results showed that the method can directly detect the virus gene in the early blood samples of the patients and complete the differential diagnosis of the flavivirus within 2 days.