Table 1.
Oligonucleotides primers used in this study.
| Purpose | Gene target | Oligonucleotide primer sequence | PCR component concentration | Thermal cycling | Amplicon size in 2% Agarose gel |
|---|---|---|---|---|---|
| Identification | ctrA | 5'ccagcggtattgtttggtggt3’ 5'caggcggcctttaataatttc3’ | 10 pmole each primer, 2.5mM MgCl2,1XPCRbuffer,200µM deoxynucleoside triphosphate, 1.25 U Taq polymerase (PerkinElmer) | 950C /4min, 35cycle 950C/10 sec,550C 30sec and1 cycle at 720C/ 7min | 177 bp |
| Serogroup characterization by multiplex PCR | Orf-2of myn B gene for Serogroup A | 5'cgcaataggtgtatatattcttcc3’ 5'cgtaatagtttcgtatgccttctt3’ | 0.3 µM each primer,5mM MgCl2, 1X PCR buffer, 200µM,1U Taq polymerase (Perkin Elmer) | 1cycle(940C/3min,550C/30sec,720C),35 cycle (920C/40sec,550C/30sec,720/20sec),1cycle (720C/10min) | 400bp |
| SiaD (serogroup B) | 5'cgtaatagtttcgtatgccttctt3’ 5'gcatgctggaggaataagcattaa3’ | 450bp | |||
| SiaD (serogroup C) | 5'gcatgctggaggaataagcattaa3’ 5'tcaaatgagtttgcgaatagaaggt3’ | 250bp | |||
| SiaD (serogroup W135) | 5'cagaaagtgagggatttccata3’ cacaaccattttcattatagttactgt3’ | 120bp | |||
| Sia D (Y) | 5'ctcaaagcgaaggctttggtta3’ ctgaagcgttttcattataattgctaa3’ | 120bp | |||