摘要：
本发明公开了一种特异性检测鲑科鱼类的实时荧光PCR试剂盒及检测方法，该试剂盒包括：PCR缓冲液、dNTP和DNA聚合酶、SEQ ID NO.1或2所示的上游引物、SEQ ID NO.3所示的下游引物、SEQ ID NO.4所示的荧光探针；其检测方法包括：1)提取待测样本DNA；2)对样本DNA和阴性对照样品进行PCR反应，并进行荧光检测；3)选择FAM荧光检测模式，基线调整取3～15个循环的荧光信号，以阈值线刚好超过正常阴性对照样品的最高点设定阈值线；若待测样品荧光增长曲线超过阈值线，并呈良好的对数增长，则判断为阳性。本发明能快速、准确地检测和鉴定鲑科鱼类成分，降低常规PCR扩增的假阳性率。
摘要（英）：
The invention discloses a real-time fluorescence PCR (polymerase chain reaction) reagent kit and a real-time fluorescence PCR detection method for specific detection of salmonidae fishes. The real-time fluorescence PCR reagent kit comprises a PCR buffer solution, a dNTP (diethyl-nitrophenyl thiophosphate) and DNA (deoxyribonucleic acid) polymerase, an upstream primer shown in the SEQ (sequence) ID (identification code) NO.1 or NO.2, a downstream primer shown in the SEQ ID NO.3, and a fluorescent probe shown in the SEQ ID NO.4. The detection method includes: (1) extracting DNA of a sample to be detected; (2) subjecting the DNA of the sample and a negative reference sample to PCR reaction, and performing fluorescence detection; (3) selecting an FAM fluorescence detection mode, adjusting a base line to take 3-15 circulating fluorescence signals, setting a threshold value line to just exceed the highest point of a normal negative reference sample; and if a fluorescence increasing curve of the sample to be detected exceeds the threshold value line and increases in excellent logarithms, judging the sample to be positive. By the aid of the reagent kit and the method, ingredients of the salmonidae fishes can be detected and identified quickly and accurately, false positive rate of normal PCR amplification is reduced.