YJAAYoung Japan Association of Arbitrators (law)
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Fifty six UPEC isolates with confirmed resistance to cefotaxime (according to CLSI, 2016) were selected to study the phylogenetic subgroups using specific primer pairs to amplify chuA, yjaA and TspE2.C4 genes.
The results of phylogenetic subgrouping of UPEC using multiplex PCR to detect chuA (279bp), yjaA (211bp) and TspE4.C2 (152bp) revealed that among 56 CRUPEC 47(83.93%) were ExPEC (37(66.07%) for B23, 6(10.72%) for B22, 3(5.36%) for D1 and 1(1.78%) for D2).
Three primer pairs used for the amplification of three genetic sequences called chuA, yjaA and TSPE4.C2 are presented in Table 1.
Sequences of oligonucleotide primers used in this study Gene Sequence Amplicon size 5'-GAC GAA CCA ACG GTC AGG AT-3' chuA 5'-TGC CGC CAG TAC CAA AGA CA-3' 279 bp 5'-TGA AGT GTC AGG AGA CGC TG-3' yjaA 5'-ATG GAG AAT GCG TTC CTC AAC-3' 211 bp 5'-GAG TAA TGT CGG GGC ATT CA-3' TSPE4.C2 5'-CGC GCC AAC AAA GTA TTA CG-3' 152 bp Gene Reference chuA (Clermont et al., 2000) yjaA (Clermont et al., 2000) TSPE4.C2 (Clermont et al., 2000) Table 2.
coli isolated from clinical samples were detected by identifying the presence of specific PCR amplified fragments (chuA, yjaA, and TspE4.C2) (Figures 1,2,3) respectively.
coli strains contained ChuA1 , YjaA , TspE4C and arp genes respectively (Fig 2).
coli is classified to four main phylogenetic groups (A, B1, B2, and D) according to the combination of two genes (chuA and yjaA) and an anonymous DNA fragment (TSPE4.C2) as described previously [22].
coli phylogenetic groups (A, B1, B2, and D) according to an established triplex PCR assay, in which the 4 phylogenetic groups yield distinct combinations of 3 possible PCR products, chuA (heme transport), yjaA (unknown function gene from E.
(26) that uses a combination of 3 DNA markers (chuA, yjaA, and an anonymous DNA fragment, TspE4.C2).