Training 16092009

Ultra2 Training at Trinity Biotech Bray, Ireland September 16th, 2009 Resolution Software Cont. Peak Labeling Error Codes 1 = Wide A peak (the A peak in the sample is 1.3 times wider) Possible causes: A peak and variant peak merging Underconcentrated FASC Overconcentrated patient samples 2 = Area of A <80% 3 = Peak area greater than expected (>5%) 4 = Peak is after A2 5 = A1c is greater than 10% (in order to confirm A1c or variant) 6 = HbF or variant is present 7 = peak area too small / big Consequences: Total area of the chromatogram is not within specs Will not be reflexed to HR Will not be transmitted to LIS 8 = %A2 is not within the normal range Hemoglobin F rules • Normal range is less than 2% • Specify the minimum %F to be labeled 0.5 – 5.0% • There are 2 RRT windows for F, F>10% and F<10% • If 14%<%F<35% the peak will be labeled with Code 6 Otherwise it will be “labeled consistent with F” • Acetyl F will be labeled if 10% or more of %F Normally it is 10 – 20% of F Hemoglobin A rules • If a peak in the A window is 15% or greater it will be labeled as A • If <15% Code 3 • If <5% will be ignored • If <80% Code 2 Hemoglobin A2 rules • Normal range 1.5 – 3.0% • If >0.1% and <10% labeled A2 • If >10% Code 3 • If less than or greater than expected Code 8 Hemoglobin A1c rules • 1 – 10% labeled A1c

Ultra2 Training at Trinity Biotech Bray, Ireland September 16th, 2009 • •

If >10% Code 5 A1c will only be labeled if A2 is also labeled

Genesys Software Beta version (has not been released yet) Only differences will be noted Setup – Assay There’s an option to print QS/HR side-by-side report after reflex. It only prints once after reflex testing. Report cannot be regenerated. Exception report can be generated. The set-up of the micro-well titer plate one can indicate which positions to dilute or skip in the option “Enter Dilution Template”. Red wells will not be sampled. Yellow wells will not be diluted e.g. hemolysate control samples. One can use either traditional round bottom plates or tall micro-well plates. Sampling will start at position A-1. Reminder: The set-up of the micro-well titer plate is completely customizable. You can use tall micro-well titer plate for adult samples (5µ L EDTA blood and fill it up to 1 mL) and run HR (the 2 min. QS on Genesys doesn’t identify/quantify A2 and F). If the laboratory is utilizing an automatic puncher analyzer, there’s a software program that converts this information and import it into Genesys with the “Import” function on the “Edit Sample List” option. It can accept multiples of the same sample identification. Additionally, there’s an option to utilize it as a diluter. Despite what’s been defined in the sample list, the U2 will honor the template set-up. The Genesys can provide phenotype labeling s.a. FA or FAU. Anything can be user-definable e.g. >20% HbA to flag whether the neonate had been transfused. Normally the percentage HbA for a baby should be 5 – 15%. Absence of HbA and only Hbvar would indicate homozygous Hbvar phenotype. Only HbS one can not differentiate HbSS or HbS-beta-thal. If HbS > HbA then sickle cell trait with beta-thal. Genesys will also label the variant - Hb Bart’s (indicator of alpha-thal). Column usage is around 700 injections.

Ultra2 Training at Trinity Biotech Bray, Ireland September 16th, 2009 Genesys.ini file AreaReject1 or 2 (is percentage) indicates which peaks will be integrated. e.g: AreaReject2=0.005 peaks < 0.5% will not be integrated after 1.4 min. Always have a copy of the ini.file on hand. Hemoglobin Genetics HbA α 2β 2 HbA2 α 2δ 2 HbF α 2γ 2 α -thal – deletion of alpha-genes then A2 below 1.5 (ref.ranges: 1.7 – 3.5%) β -thal – A2 above 3.5% β -thal combined with β -variant then beta-variant is elevated Ex: β -thal combined with HbS(β -variant) then HbS(beta-variant) is elevated (60-70%) β -thal combined with α -variant then alpha-variant is decreased α -thal combined with β -variant then beta-variant is decreased Ex: α -thal combined with HbS(β -variant) then HbS(beta-variant) is decreased (2530%) α -thal combined with α -variant then alpha-variant is elevated HbH Hb Bart’s

β 4 γ 4