It is a ppt about is HPLC should be used as routine screening test for anemic and antenatal cases.This ppt is prepared by taking information from journal

1. MICROCYTIC HYPOCHROMIC
ANEMIA:SHOULD HPLC BE USED
ROUTINELY FOR SCREENING
ANEMIC & ANTENATAL PATIENTS?

2. INTRODUCTION:
 Hemoglobin mutations form the most common human single disorders
 About 698 genetically different hemoglobin variants are found scattered all over the world causing various
hemoglobinopathies.
 About 5.2% of the world population (and more than 7% of pregnant females) carry a significant variant.
 . About 1.1% of couples around the world are at risk for having children with a hemoglobin disorder of which 2.7
per 1000 conceptions are actually affected.
 Hemoglobin disorders contribute to 3.4% of mortality in children aged less than five years worldwide.
 Among these disorders, sickle cell syndromes and thalassemia constitute major public health problems.
 The frequency of beta-thalassemia trait in India has been reported to vary from <1 to 17% depending on the
region studied, with an average of 3.3%.
 The average frequency of sickle cell disease is 4.3%.
 Hemoglobin E has been reported as the most common hemoglobin (Hb) variant in Southeast Asia and the
second most prevalent worldwide.
 The clinical manifestations of these hemoglobinopathies can vary from asymptomatic states to severe, lifelong,
transfusion-dependent anemia with multi organ involvement and severely reduced life expectancy.

3. AIM & OBJECTIVES
 To determine the prevalence of hemoglobinopathies in patients with microcytic hypochromic
anemia and to assess the suitability of using high performance liquid chromatography (HPLC)
routinely for screening antenatal cases and patients with anemia.

4. MATERIALS & METHODS
- A total of 4335 cases received from Mar 2007 - Nov 2011 for hemoglobin (Hb) variant analysis were studied for
various hemoglobinopathies and variants. The tests were performed on BIO RAD ‘VARIANT’.
- The instrument utilizes the principle of high performanc liquid chromatography (HPLC).
- Patients included cases of microcytic hypochromic anemia, referred to this centre where a co-existent
hemoglobinopathy was suspected, while other cases were of anemia with no apparent underlying cause,
antenatal cases, transfusion requiring children and adults and their family members.
- About 2 ml of blood sample was collected in EDTA vial. They were stored at 4-8°C and were analyzed in batches
once a week.
- Patients referred to us either already had a hemogram and a peripheral blood smear (PBS) reported or it was
concurrently asked for.
- Concurrent iron deficiency anemia (IDA) was ruled out on the basis of iron studies including serum iron
levels, total iron binding capacity, transferrin saturation and serum ferritin levels.
- Samples with abnormal results were also run on KX-21 hematologyanalyzer to obtain hemoglobin values and
indices.

5. - Sickling test was performed using freshly prepared sodium meta bisulphite when S-window was
eluted in the sample.
- Other confirmatory tests including acid and alkaline electrophoresis were done when required.

8. RESULTS