|Test Code||Test||Specimen Requirement|
|B5021||HPLC ANALYSIS OF HAEMOGLOBINS
Analysis of different haemoglobins and their relative quantitation by cation exchange HPLC with BIORAD Variant-II Hb analysis system using β-Thal Short program.
|2mL of EDTA anticoagulated peripheral blood (Purple top)- 2 tubes. Stored at 4°C.|
|B5024|| Mutation analysis:
Thalassaemia mutation analysis includes the following comprehensive workflow done as required by the requests, clinical conditions, Hb Variant results and other lab findings.
I. Screening of nine common mutations in globin gene by Reverse Dot Blot method:
a) Codon 8/9(+G) Mutation.
b) Codon 15(G>A) Mutation.
c) IVS-I-1(G>T) Mutation.
d) Codon 30 (G>C) Mutation.
e) IVS-I-5(G>C) Mutation.
f) IVS-I-1(G>A) Mutation.
g) Codon 41/42 (-TCTT) Mutation.
h) Codon 26(G>A) (βE) Mutation.
i) Codon 6 (A>T) (βS) Mutation.
II. Screening of rare mutations in the β Globin gene
1. Screening of 619bp deletion by PCR.
2. Gene dosage analysis in β Thalassaemia by capillary electrophoresis.
3. Screening of δβ (Asian-Indian deletion) thalassaemia by GAP PCR.
4. Screening Hb LEPORE mutation causing δβ gene fusion by GAP PCR.
5. Screening of HPFH3 deletion by GAP PCR.
6. Screening of rare deletions / duplications by MLPA analysis (β)
(Multiplex Ligation-dependent Probe Amplification)
7. Analysis of -158 Gγ (C>T) XmnI polymorphism by RFLP method.
8. Screening for other point mutations, small insertions and deletions causing β Thalassaemia by Sanger’s DNA Sequencing.
9. Molecular characterization of other abnormal β-globin variants by DNA sequencing.
III. Screening of eight Common deletions causing β Thalassaemia by Multiplex-PCR and agarose gel electrophoresis
1. α3.7 deletion.
2. -α4.2 deletion.
3. --SEA deletion.
4. --THAI deletion.
5. --20.5 deletion.
6. --MED deletion.
7. --FIL deletion.
8. --SA deletion.
IV. Screening of rare mutations in α Thalassaemia:
1. Gene dosage analysis by multiplex PCR followed by capillary electrophoresis.
2. Poly-A deletion screening in α2 gene by gene scan.
3. Screening of other point mutations, small insertions and deletions in α1 and α2 genes by Sanger’s DNA sequencing.
4. Screening of rare deletions / duplications by MLPA analysis (α) (Multiplex Ligation-dependent Probe Amplification)
5. Molecular characterization of other abnormal α globin variants by DNA sequencing.
|9 mL of EDTA anticoagulated peripheral blood (Purple top) tube. Stored at 4°C.|
|B5025,B5092||Prenatal diagnosis for β-Thalassaemia:
1. Screening of carrier status of the propositus and husband (with Thalassaemia mutation analysis workup)
2. Screening of such identified mutations in the foetal DNA (Chorionic villus sample testing)
|9 mL of EDTA anticoagulated peripheral blood (purple top) tube. Stored at 4°C.|