公开(公告)号：US20050170341A1

公开(公告)日：2005-08-04

申请号：US10138162

申请日：2002-05-02

申请人： Roger Stich ,  Yasuko Rikihisa

发明人： Roger Stich ,  Yasuko Rikihisa

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/689

摘要： Tools and methods for detecting the presence of E. canis and E. chaffeensis in a sample obtained from an animal are provided. The methods employ a polymerase chain reaction and primer sets that are based on the p30 gene of E. canis and the p28 gene of E. chaffeensis. The present invention also relates to the p30 and the p28 primer sets. Each p30 primer set comprises a first primer and the second primer, both of which are from 15 to 35 nucleotides in length. The first p30 primer comprises a sequence which is complementary to a consecutive sequence, within the following sequence: CCA AGTGTCTCAC ATTTTGGTAG CTTCTCAGCT AAAGAAGAAA GCAAATCAAC TGTTGGAGTTTTTGGATTAA AACATGATTG GGATGGAAGT CCAATACTTA AGAATAAACA CGCTGACTTTACTGTTCCAA AC. SEQ ID NO.1. The second p30 primer comprises a sequence which is complementary to the inverse complement of a consecutive sequence contained within the following sequence: GTTACT CAATGGGTGG CCCAAGAATA GAATTCGAAA TATCTTATGA AGCATTCGAC GTAAAAAGTC CTAATATCAA TTATCAAAAT GACGCGCACA GGTACTGCGC TCTATCTCAT CACACATCGG CAGCCAT, SEQ ID NO.2. The first p28 comprises a sequence which is complementary to a consecutive sequenc, within the following sequence: A GTTTTCATAA CAAGTGCATT GATATCACTA ATATCTTCTC TACCTGGAGT ATCATTTTCC GACCCAACAG GTAGTGGTAT TAACGG, SEQ ID NO. 3. The second p28 primercomprises a sequence which is complementary to the inverse complement of a consecutive sequencewithin one of the following two sequences: CAT TTCTAGGTTT TGCAGGAGCT ATTGGCTACT CAATGGATGG TCCAAGAATA GAGCTTGAAG TATCTTATGA, SEQ ID NO. 4, or C AAGGAAAGTT AGGTTTAAGC TACTCTATAA GCCCAGA, SEQ ID NO. 5.

摘要翻译： 提供了用于检测从动物获得的样品中E. canis和E.Chaffeensis的存在的工具和方法。 该方法采用聚合酶链反应和引物组，其基于大肠杆菌的p30基因和E.Chaffeensis的p28基因。 本发明还涉及p30和p28引物组。 每个p30引物组包含长度为15至35个核苷酸的第一引物和第二引物。 第一个p30引物包含与以下序列中的连续序列互补的序列：CCA AGTGTCTCAC ATTTTGGTAG CTTCTCAGCT AAAGAAGAAA GCAAATCAAC TGTTGGAGTTTTTGGATTAA AACATGATTG GGATGGAAGT CCAATACTTA AGAATAAACA CGCTGACTTTACTGTTCCAA AC。 SEQ ID NO.1。 第二个p30引物包含与以下序列中包含的连续序列的反向互补序列互补的序列：GTTACT CAATGGGTGG CCCAAGAATA GAATTCGAAA TATCTTATGA AGCATTCGAC GTAAAAAGTC CTAATATCAA TTATCAAAAT GACGCGCACA GGTACTGCGC TCTATCTCAT CACACATCGG CAGCCAT，SEQ ID NO.2。 第一个p28包含与以下序列中的连续序列互补的序列：GTTTTCATAA CAAGTGCATT GATATCACTA ATATCTTCTC TACCTGGAGT ATCATTTTCC GACCCAACAG GTAGTGGTAT TAACGG，SEQ ID NO： 第二个p28引物包含与以下两个序列之一的连续序列的反向互补序列互补的序列：CAT TTCTAGGTTT TGCAGGAGCT ATTGGCTACT CAATGGATGG TCCAAGAATA GAGCTTGAAG TATCTTATGA，SEQ ID NO： 4或C AAGGAAAGTT AGGTTTAAGC TACTCTATAA GCCCAGA，SEQ ID NO： 5。