As rb presentation

1. Prevalence of Hereditary Hemochromatosis
(HEF) gene mutations and its effect on serum
ferritin and serum iron levels.
Name: Dr. Sikandar Ali Khan
Supervisor Name: Dr. Yasar Mehmood Yousafzai
Co-Supervisor Name: Dr. Sami Siraj
Institute/Department: Institute of Basic Medical Sciences,
Khyber Medical University

2. Introduction
• Hereditary Hemochromatosis is characterized by abnormal synthesis of Human
Hemochromatosis Protein, resulting in increase serum iron and serum ferritin.
• One of the common single gene disorders in the western world.
• The exact prevalence of classic hereditary hemochromatosis is unknown. In
individuals of Northern European descent the prevalence is estimated to be as
high as 1 in 227 individuals in the general population.
• Hereditary Hemochromatosis Gene Mutations has a reported prevalence of 8% in
British Pakistani people, in one study.

3. Rationale
• No studies have been undertaken to determine the prevalence/frequency
of Hereditary Haemochromatosis Gene Mutations in Peshawar district.
• Hereditary Haemochromatosis Gene mutations leads to certain clinical
conditions which can be ranging from minor to lethal.
• Therefore, true prevalence of HFE Gene Mutation must be determined in
order to develop a reliable screening protocol and molecular diagnosis for
these patients.

4. Objectives
• To determine the prevalence of Hereditary Hemochromatosis Gene
mutations among the general population of Peshawar District.
• To correlate the Hereditary Hemochromatosis Gene mutations with
hemoglobin levels, RBC count, MCV, MCH, MCHC, RDW, serum iron and
serum Ferritin levels.

5. Operational Definitions
• Iron Overload:
• In men normal values of serum ferritin are from 12 to 300ng/ml in blood
and in females it ranges from 12 to 150 ng/ml. The results may vary
slightly among laboratories.
• If the values are above these normal limits, it is called increase iron, but if
the values are above 1000ng/ml, it is called iron overload.
• HFE gene:
• Human hemochromatosis protein also known as the HFE protein is a
protein which in humans are encoded by HFE gene. The HFE gene is
located on short arm of chromosome 6 at location 6p21.3.

6. Operational Definitions
• ANEMIA: Anemia will be defined as hemoglobin values of
 Less than 13 g/dL in men and
 Less than 12 g/dL in women.
 Less than 11.0 –11.5 g/dL in children
(WHO)
• MICROCYTOSIS: Microcytosis will be defined as MCV less than 80fL.
• HYPOCHROMIA: Hypochromia will be defined as MCH less than 27pg.

7. Methodology & Analysis Plan
• Study Design: observational cross-sectional study.
Settings: (i) District Peshawar,
(ii) IBMS KMU Peshawar.
• Duration: 6 months
• Sample Size: Keeping margin of error at 5%, confidence level 95% and
population of Peshawar at 4.2 million, prevalence of 8% the sample size
was calculated to be 364.
• Sampling Technique: Multi-stage randomized cluster sampling

8. Methodology & Analysis Plan
• Sample Selection
• Inclusion criteria:
 5 – 62 years
 Both genders
• Exclusion Criteria:
Not available in the household.
Had iron supplementation in past three months.
Had Blood transfusion in past three months.

9. Methodology & Analysis Plan
Participants from local UCs
of Peshawar
•Informed
Consent
Sample
Collection
•5 cc blood in EDTA
tubes
CBC and
Peripheral
Smear
•MCV,MCH,
MCHC,
•RDW
Anemic
Not
Anemic
Extraction of
Genomic DNA
PCR
•HFE gene
Mutation
Gel Electrophoresis

10. Methodology & Analysis Plan
• Data will be collected after allocating unique study number to each patient.
• Data will be put in Microsoft excel and exported to SPSS for analysis.
• Numerical variables will be presented by mean ±SD.
• Categorical variable i.e. gender will be presented by frequency and
percentage.
• Proportional symbols maps will be created for prevalence of mutation in
HFE Gene in the population units of district Peshawar.
• Correlation will be calculated for HFE Gene mutations with hemoglobin
levels, RBC count MCV and MCH, MCHC, RDW, serum iron and serum
Ferritin.

11. References:
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• 3. Katsarou MS, Latsi R, Papasavva M, Demertzis N, Kalogridis T, Tsatsakis AM, et al. Population-based analysis of the
frequency of HFE gene polymorphisms: Correlation with the susceptibility to develop hereditary hemochromatosis.
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• 6. Feder JN, Penny DM, Irrinki A, Lee VK, LEBRon JA, Watson N, et al. The hemochromatosis gene product complexes
with the transferrin receptor and lowers its affinity for ligand binding. Proceedings of the National Academy of Sciences.
1998;95(4):1472-7.
• 7. Bring P, Partovi N, Ford JAE, Yoshida EM. Iron overload disorders: treatment options for patients refractory to or
intolerant of phlebotomy. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy. 2008;28(3):331-42.
• 8. Elmrghni S, Dixon RA, Williams DR. Frequencies of HFE gene mutations associated with haemochromatosis in the
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• 9. Weinberg ED. Exposing the hidden dangers of iron: what every medical professional should know about the
impact of iron on the disease process: Cumberland House Publishing; 2004.

12. Thank You
Any Questions