逆转录聚合酶链式反应快速检测水稻条叶枯病毒  

Detection of Rice Stripe Virus by Reverse Transcription and Polymerase Chain Reaction

作　　者：吕英芝 邓可京 曲志才 严键 沈大棱 

出　　处：《复旦学报（自然科学版）》1999年第5期591-593,共3页Journal of Fudan University：Natural Science

基　　金：美国McKnight基金;上海市科委资助

摘　　要：逆转录聚合酶链式反应（Ｒｅｖｅｒｓｅ Ｔｒａｎｓｃｒｉｐｔｉｏｎ ａｎｄ Ｐｏｌｙｍ ｅｒａｓｅ Ｃｈａｉｎ Ｒｅａｃｔｉｏｎ，ＲＴ－ＰＣＲ）具有灵敏度高、准确性好的特点，因而被广泛应用于植物病毒检测．在ＲＴ反应中，ＲＮＡ 样品的快速制备尤为重要．用异丙醇二步沉淀法获得ＲＮＡ，快速检测水稻叶片中ＲＳｔＶ 的ＲＴ－ＰＣＲ 方法，可使测定时间缩短至６ ｈ，并可从０．０７８ｍ ｇ 感病叶片中检测出ＲＳｔＶ．A pair of primers is designed and synthesized based on the nucleotide sequence of coat protein(CP) gene of rice stripe virus (RStV) genome RNA3 and used to amplify a 980 bp DNA fragment by reverse transcription and polymerase chain reaction (RT PCR) to detect RStV from RStV infected rice leaves. The precipitation method using isopropanol shortens the diagnostic process and is very suitable for simultaneous RNA and DNA isolation from very small amount of material. It took only 6 h to finish detection of the virus. RStV RNA can be detected in an RNA extract as little as 0.078 mg of infected rice leaves. It shows that the RNA sample extracted by isopropanol precipitation method can be used as template for reverse transcription procedure.

关 键 词：逆转录 聚合酶 链式反应 水稻条叶枯病毒 

分 类 号：S432.41[农业科学—植物病理学] S435.121.9[农业科学—农业昆虫与害虫防治]