Four partial fragments from cox1, rrnS, cob and cox2 gene regions were PCR-amplified for B. xylophilus using universal primer sets (LCO1490/HCO2198, 12SL1091/12SH1478 and CtybL14841/CtybH15149 for cox1, rrnS and cob, respectively) and the primer set (COX2-F/R) designed directly from conserved regions of cox2 sequences for chromadorean nematode sequences (See Additional file 6 for primer sequences).
In addition, HA and NA genomic fragments were amplified from the patient's respiratory tract swab and the samples from the poultry booths and wholesale market using universal primer sets as described previously, and subjected to sequencing.
For P. vulnus, two partial fragments (from cob and cox2 gene regions) were amplified using universal primer set CtybL14841/CtybH15149 for cob and the primer set (COX2-F/R).
Primers and cycling conditions are listed in Table 2. Specificity of MSP primers in detecting the DKK3 methylation status were demonstrated using universal unmethylated and universal polymethylated DNA as a template (Epi Tect Control DNA Set; Qiagen, Hilden, Germany).
Using universal eukaryotic primer sets (Table S1), nearly full-length 18S ribosomal RNA (rRNA) genes were amplified from the genomic DNA.
Initially, partial mtDNA fragments were amplified from different regions of mtDNA using universal primers or primer sets designed from the conserved regions of closely related nematode species.
PCR amplification of the coding region of the 8 influenza virus genes in the cDNA was performed as described (7 ) by using universal gene-specific primer sets (3 ).
The total OATP1B3 mRNA (Ct- plus Lt-OATP1B3) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA levels were respectively determined using universal probe #59 with the primer set (5'-CGGCCTandCTTGACCTATG-'3 and 5'-TGAGTTGCAATAAGAAAGTGGTACA-3'), and probe #60 with the primer set (5'-andCACATCGCTCAGACAC-'3 and 5'-GCCCAATACGACCAAATCC-3').
Partial fragments from different regions of the mitochondrial genome were initially amplified and directly sequenced using universal primer combinations or primer sets designed directly from the conserved regions of nematode mitochondrial gene sequences ([ 51]; see Additional file 4 for primer details).
We determined the incidence, complications and risk factors for the development of HZ after renal transplantation in a setting using universal antiviral prophylaxis.
