Original article - Blood Transfusion

More documents

Recommendations

Info

HPA genotyping in southern ItalyHPA-1: Nci I (1U) 37°C over night → HPA-1a: 247bpHPA-1b: 77+170bpHPA-1aHPA-1b247bp↓Nci 1↓CTGGG247bp77bp CCGGG 170bp↑Nci 1HPA-3: Fok I (1U) 37°C over night → HPA-3a: 126+127 bpHPA-3b: 253 bpHPA-3aHPA-3b253bp↓Fok 1↓126bp CATCC 127bp↑Fok I253bpCAGCCHPA-5: Mnl I (1U) 37°C over night → HPA 5a: 8+33+136+97 bpHPA 5b: 8+169+97 bpHPA-5aHPA-5b274bp↓Mnl 1↓8bp 33bp 136bp GAGG 97bp↑ ↑ ↑Mnl I Mnl I Mnl I8bp 169bp AAGG 97bp↑↑Mnl IMnl IFigure 1 - Diagrammatic representation of the restriction site map of the PCR amplified fragments of HPA-1, HPA-3, HPA-5 generatedafter digestion. The arrows indicate the positions of the restriction enzyme sitestable I. PCR was performed according to the protocolsummarised in tables II and III. The position of the enzymerestriction sites on the bp PCR product and the length ofthe fragment generated after digestion by Nci I (HPA-1),Fok I (HPA-3) and Mnl I (HPA- 5) are schematically shownin figure 1. After amplification, the PCR products weredigested with restriction enzymes under the conditionssummarised in table IV. The restriction products weresubmitted to gel electrophoresis in a 3% agarose gel for120’ at 100-150 Volt in TAE buffer. DNA fragments wereresponsabile delle gravi forme di FNAITP osservate nei 12neonati ed è stata confermata dalla presenza di anticorpilesivi nel siero materno (10 anti-HPA-1a e 2 anti-HPA-1b).La figura 2 mostra un esempio delle analisi PCR-SSPper i 6 sistemi HPA indagati.DiscussioneCiascun sistema HPA consiste di 2 varianti alleliche,<strong>Blood</strong> Transfus 2005; 3: 222-30225
G Fratellanza et al.Table V - HPA genotype and antigens frequencies in the studied populationHPA Genotype Number of subjects Genotype Frequencies (%) HPA Antigens Antigen Frequency (%)Gene FrequencyHPA-1 a/a 144 72% HPA-1 a 95.5% 0.838HPA-1 a/b 47 23.5%HPA-1 b/b 9 4.5% HPA-1 b" 28% 0.162HPA-2 a/a 146 73% HPA-2 a 97% 0.850HPA-2 a/b 48 24%HPA-2 b/b 6 3% HPA-2 b 26.5% 0.150HPA-3 a/a 92 46% HPA-3 a 85.5% 0.658HPA-3 a/b 79 39.5%HPA-3 b/b 29 14.5% HPA-3 b 54% 0.342HPA-4 a/a 199 99.5% HPA-4 a 100% 0.998HPA-4 a/b 1 0.5%HPA-4 b/b 0 0% HPA-4 b 0.5% 0.002HPA-5 a/a 169 84.5% HPA-5 a 99.5% 0.920HPA-5 a/b 30 15%HPA-5 b/b 1 0.5% HPA-5 b 15.5% 0.080HPA-6 a/a 199 99.5% HPA-6 a 100% 0.998HPA-6 a/b 1 0.5%HPA-6 b/b 0 0% HPA-6 b 0.5% 0.002Figure 2 - PCR-SSP agarose gel determination of HPA polymorphisms in six donors226<strong>Blood</strong> Transfus 2005; 3: 222-30
Page 3: G Fratellanza et al.Table I - Seque
Page 7 and 8: G Fratellanza et al.contrast, the f
Page 9: G Fratellanza et al.References1) Kr

Original article - Blood Transfusion

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?