Fig_1.tif (1.09 MB)
Download file

Multiplex-PCR Assay For Helicobacter-specific Virulence Factors.

Download (0 kB)
figure
posted on 19.08.2015, 02:52 by Sinem Oktem-Okullu, Arzu Tiftikci, Murat Saruc, Bahattin Cicek, Eser Vardareli, Nurdan Tozun, Tanil Kocagoz, Ugur Sezerman, Ahmet Sinan Yavuz, Ayca Sayi-Yazgan

(a) amplification of virulence genes of H. pylori G27 strain by multiplex PCR; Lane M, 100 bp- marker (ThermoSCIENTIFIC, Gene Ruler), Lane 1 (Reaction 1); ureA (244bp), ureB (645bp), hpaA (534bp), cagA (415bp), napA (384bp), Lane 2 (Reaction 2); dupA (584bp), oipA (401bp), vacA (333bp), and Lane 3 (Reaction 3); babA (832bp) (b) PCR inferred results of s1/s2 and m1/m2 allelles of vacA gene; Lane M, 100 bp- marker (ThermoSCIENTIFIC, Gene Ruler), Lane 1 (reaction 1); vacA s1 (259bp), Lane 2 (Reaction 2); vacA s2 (286 bp), Lane 3 (Reaction 3); vacA m1 (567 bp), and Lane 4 (Reaction 4) vacA m2(642 bp). (c) Multiplex-PCR application of the biopsy samples taken from randomly selected patients with ulcer and gastritis; Lane M, 100 bp- marker (ThermoSCIENTIFIC, Gene Ruler), between Lane 1 to 6 are representing patients with gastritis or ulcer: Multiplex PCR reaction results of patient 1 and 2 (gastritis or ulcer) are shown between Lane 1 to 3 and Lane 4 to 6, respectively. Reaction 1 (Lane 1 and 4) ureA (244bp), ureB (645bp), hpaA (534bp), cagA (415bp), napA (384bp), Reaction 2 (Lane 2 and 5); dupA (584bp), oipA (401bp), vacA (333bp), and Reaction 3 (Lane 3 and 6); babA(832bp) (d) PCR inferred results of s1/s2 and m1/m2 allelles of vacA gene of randomly selected patients with gastritis and ulcer respectively; Lane M, 100 bp- marker (ThermoSCIENTIFIC, Gene Ruler), between Lane 1 to 3 are representing patients with gastritis (left side) or ulcer (right side): Patients with gastritis; Lane 1–3, vacA s1 (259bp), and vacA s2 (286 bp), patients with ulcer; Lane 1–3, vacA m1 (567 bp), and vacA m2 (642 bp). (e) Multiplex urease-PCR assay to detect the Helicobacter positive and negative samples; Lane M, 100 bp- marker (ThermoSCIENTIFIC, Gene Ruler), Lane 1–2 are from randomly selected patients, and lane 3 is from H.pylori positive control strain G27. Sybr Gold (Invitrogene) was used for the gel in (a) and (c) and gel pictures were taken by using Observable Real Time Gel Electrophoresis System (Salubris Technica, Turkey).

History