Human Infectiology MDx | Catalogue 2011 - Diagenode Diagnostics

HUMAN MOLECULAR DIAGNOSTICS CATALOGUE 2011GASTROENTERITIS INFECTIONSShigellaRef: Dia-Shigella-050Shigella is a genus of Gram-negative, non-spore formingrod-shaped bacteria closely related to Escherichia coli andSalmonella.Shigella species are classified by four serogroups:• Serogroup A: S. dysenteriae (12 serotypes)• Serogroup B: S. flexneri (6 serotypes)• Serogroup C: S. boydii (23 serotypes)• Serogroup D: S. sonnei (1 serotype)Shigella causes dysentery that results in the destruction of theepithelial cells of the intestinal mucosa in the cecum and rectum.Some strains produce enterotoxin and Shiga toxin, similar to theverotoxin of E. coli O157:H7 (1) . Both Shiga toxin and verotoxin areassociated with causing hemolytic uremic syndrome.(1)Hale TL, Keusch GT (1996). Shigella. in: Baron’s Medical Microbiology (BaronS et al., eds.) (4th ed.). Univ of Texas Medical Branch.VALIDATIONResearched pathogen:• Shigella: FAM dye, emission: 520 nmTarget genes:• ipaH (Shigella)Number of wells by test: 1 (Shigella)Samples used:• StoolValidated extraction methods: • Nuclisens easyMAG ® System (Biomerieux)• QIAamp DNA Stool mini kit (Qiagen)• MagNa pure LC system (Roche)• QiaSymphony (Qiagen)Real-time PCR systems: • ABI (7000/7300/7500/7900)• Roche lightCycler (2.0/480)• Biorad (ICycler/IQ5/CFX96)• Qiagen RotorGene (6000)• Stratagene (MX3000P/3005P)• Cepheid (SmartCycler II)PCR volumes: 50 µl, 25 µl, 20 µl (depending of the real-time PCR system used)PCR reactions/kit:• 50 (50 µl PCR)• 100 (25 µl PCR)• 120 (20 µl PCR)In combination with:• Universal inhibition control (DIA-UIC-050):DIA-UIC(YD)-050: yellow dye, emission: 549 nmDIA-UIC(DR)-050: orange dye, emission: 576 nmDIA-UIC(Cy5)-050: Cy5 dye, emission: 662 nm• DNA extraction & PCR inhibition control (DIA-EIC/DNA)-050:DIA-EIC/DNA(YD)-050: yellow dye, emission: 549 nmDIA-EIC/DNA(DR)-050: orange dye, emission: 576 nmDIA-EIC/DNA(Cy5)-050: Cy5 dye, emission: 662 nm10 | www.diagenodediagnostics.com