International Journal of Current Research and Review
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IJCRR - 8(17), September, 2016

Pages: 46-49

Date of Publication: 11-Sep-2016


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STUDY OF LABORATORY PARAMETERS IN HEMOPHILIA PATIENTS

Author: T. B. Sadaria, H. M. Goswami, Safal Patel

Category: Healthcare

Abstract:Introduction: Haemophilia is X- linked congenital bleeding disorder with a frequency of about one in 10,000 births. Haemophilia is caused by deficiency of coagulation factor VIII (haemophilia A) or factor IX (haemophilia B) related to mutations of clotting factor gene. Objectives: To check effectiveness of various screening and confirmatory tests for diagnosis of haemophilia. Methods: Retrospective study of laboratory diagnosis of haemophilia was conducted in our haematology section of pathology department of Tertiary Care Teaching Centre from 1 August 2014 to 30 July 2016. Patients in the age group of 0 year to 55 years with factor VIII and factor IX level below 50% of normal were included in the study. Routine haematological tests like haemoglobin and platelet count and coagulation profile of patient for prothrombin time, activated partial prothrombin time, factor VIII and factor IX level were analysed. Results: Out of 122 cases, 97 cases were of haemophilia A while 25 cases were of haemophilia B. Haemoglobin count of patients ranged from6gm% to 14.2 gm%. Platelet count and PT (prothrombin time)of patients were within normal limits. APTT (Activated Partial Thromboplastin Time) was prolonged (41.6 sec. to 124 sec) in all patients. Factor VIII level was reduced (< 50%) in all patient of haemophilia A. Factor IX level was reduced (< 50%) in all patient of haemophilia B. Conclusion: Laboratory analysis of blood for haemoglobin, platelets, PT and APTT help to suspect the diagnosis. Diagnosis of haemophilia is confirmed by factor VIII assay for haemophilia A and factor IX assay for haemophilia B. Level of factor VII and IX also decide severity of disorder.

Keywords: Haemophilia A, Haemophilia B, Factor VIII, Factor IX

Full Text:

Introduction:
Haemophilia is X- linked congenital bleeding disorder with a frequency of about one in 10,000 births. Haemophilia is caused by deficiency of coagulation factor VIII (haemophilia A) or factor IX (haemophilia B) related to mutations of clotting factor gene.
Objectives: To check effectiveness of various screening and confirmatory tests for diagnosis of haemophilia.
Methods: Retrospective study of laboratory diagnosis of haemophilia was conducted in our haematology section of pathology department of Tertiary Care Teaching Centre from 1 August 2014 to 30 July 2016. Patients in the age group of 0 year to 55 years with factor VIII and factor IX level below 50% of normal were included in the study. Routine haematological tests like haemoglobin and platelet count and coagulation profile of patient for prothrombin time, activated partial prothrombin time, factor VIII and factor IX level were analysed.
Results: Out of 122 cases, 97 cases were of haemophilia A while 25 cases were of haemophilia B. Haemoglobin count of patients ranged from6gm% to 14.2 gm%. Platelet count and PT (prothrombin time)of patients were within normal limits. APTT (Activated Partial Thromboplastin Time) was prolonged (41.6 sec. to 124 sec) in all patients. Factor VIII level was reduced (<50%) in all patient of haemophilia A. Factor IX level was reduced (<50%) in all patient of haemophilia B.
Conclusion: Laboratory analysis of blood for haemoglobin, platelets, PT and APTT help to suspect the diagnosis. Diagnosis of haemophilia is confirmed by factor VIII assay for haemophilia A and factor IX assay for haemophilia B. Level of factor VII and IX also decide severity of disorder.

AIMS AND OBJECTIVES
Objectives of the Study:
1) Approach to haemophilia with detailed clinical evaluation and laboratory diagnosis.
2) To evaluate various screening and confirmatory tests for diagnosis of haemophilia.
3) To effectively treat patient with haemophilia according to various laboratory parameter.

MATERIAL AND METHODS
Retrospective study of laboratory diagnosis of haemophilia was conducted in our haematology section of pathology department of Tertiary Care Teaching Centre for the periods of two years from 1 August 2014 to 30 July 2016. All patients referred to haematology section of pathology department in the age group of 0 year to 55 years with factor VIII and factor IX level below 50% of normal were included in the study. Routine haematological tests like haemoglobin and platelet count were analysed. Coagulation profile of patient for prothrombin time, activated partial prothrombin time, factor VIII and factor IX level were also considered. EDTA anti -coagulated fresh whole blood sample were obtained for Haemoglobin and Platelet count. They were done in CELL-dyn 3700 haematology analyser. Haemoglobin is measured by modified hemiglobinhydroxylamine or modified hemiglobincyanide method. Platelet count was confirmed by manual method. Samples for coagulation tests were collected in vacuette containing 0.2 ml 3.2%sodium citrate solution and centrifuged for 15 minutes at 4000 rpm. These samples were analysed in Diagnostica Stago fully automated coagulometer. Reagents used for Prothrombin Time were NEOLASTIN CI PLUS, STA CLEANER SOLUTION, STA DESORB U.Reagents used for Activated Partial Thromboplastin Time were C.K. Prest, Cacl2 0.025 m, STACLEANER SOLUTION. Reagents used for Factor VIII were STA-Deficient Factor VIII Kit- lyophilized citrated human plasma from which factor VIII has been removed by selective immunoadsorption and for Factor IX were STA-Deficient Factor IX Kit- lyophilized citrated human plasma from which factor IX has been removed by selective immunoadsorption.

RESULTS
“Study of Laboratory Parameters in Haemophilia Patients” was a retrospective study of 122 patients of haemophilia, conducted over a period of two years. Out of 122 cases, 97 cases were of haemophilia A while 25 cases were of haemophilia B. Haemophilia A was more common than haemophilia B. Age range of these patients very from 0-55 years. Majority of patients were below age of 20 years. Haemoglobin count of patients with haemophilia A and B ranged from 6gm% to 14.2 gm%. Platelet count of patients with haemophilia A and B ranged from 1,50,000 /dl to 4,91,000/dl that was within normal limits. Prothrombin time was normal in all of these patients. It ranged from 10 sec. to 16.5 sec. Control for PT was 13.5 sec. APTT was prolonged in all patients with haemophilia A and B. It ranged from 41.6 sec. to 124 sec. Normal control of APTT was 26 sec. to 40 sec. (Table 1) Factor VIII level was reduced (<50%) in all patient of haemophilia A. Normal range for factor VIII was 50% to 150% (0.5 IU/ml to 1.5 IU/ml). According to factor VIII level, haemophilia A is divided into mild, moderate and severe. (Table 2) Factor IX level was reduced (<50%) in all patient of haemophilia B. Normal range for factor VIII was 50% to 150% (0.5 IU/ml to 1.5 IU/ml). According to factor IX level haemophilia B is divided into mild, moderate and severe. (Table 3)

DISCUSSION
Retrograde study of haemophilia involving 122 cases was done during period of two years from 1stAugust 2014 to 30th July 2016 and diagnosis was made with available clinical and laboratory data. Results were analysed and discussed with previous similar studies. In present study Haemoglobin level ranged from 6gm% to 14.2gm%. 36 cases (29%) had moderate reduction in haemoglobin (11gm%) which correlated well with study done by Steven et al5 showing 25%of cases. This decrease in haemoglobin is explained by loss of blood due to repeated bleeding episodes. Platelet count of all patients was ranging from 1,50,000 to 4,91,000 /cmm which correlated well with study done by Agarwal et al.6 Prothrombin time of all patients was between 11-16.5 second that is within normal limits. This correlated well with study done by Kitchens CS.7 APTT was prolonged in all cases of haemophilia A and B. In majority of patients, APTT was more than 60 seconds. These findings correlated well with study done by Kitchens CS7 and Takim and Shrivastava.8 Most of patients with APTT more than 80 seconds belong to severe haemophilia. This support the fact that if APTT is twice the control, chances are that patients have severe haemophilia. (Graph 1 and 2)

Factor assay was done in all cases; findings correlate with study done by Shanthala Devi et al9 and Rodgers gm and Charles SG.10 Most of cases of haemophilia were of severe degree. (Graph 3 and 4)

CONCLUSION
• Haemophilia A is more common than haemophilia B.

• Both haemophilia A and B are more commonly seen in male below 20 years of age.

• Laboratory analysis of blood for haemoglobin, platelets, PT and APTT help to suspect the diagnosis along with clinical findings but not confirmatory.

• Diagnosis of haemophilia is confirmed by factor VIII assay for haemophilia A and factor IX assay for haemophilia B. Level of factor VII and IX also decide severity of disorder

ACKNOWLEDGEMENT
The author acknowledges the help received from Dr. Hansa M. Goswami, M.D. PATH., Professor and Head, Department of Pathology for teaching me the scientific approach of the subject and its subtle aspects. I would like to give my special thanks to all the technicians of haematology section, Department of Pathology, B.J. Medical College, Ahmedabad for helping me while conducting study. Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. Authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.

References:

1. Shrivastava A. Haemophilia in developing countrieschallenge of detection and diagnosis. In Sohail MT, Heijunen L. comprehensive haemophilia case in developing countries. Lahore : Ferozsons. 2001: p.17-25

2. Peter J. Guidelines for development of a national programme for haemophilia. Geneva: world health organization.1996.1-76.

3. Levine PH, Brettler DB. Clinical aspects and therapy of haemophilia A. In Ronald H. editor. HaematologyBasic Principle and Practice. New York: Churchill Livingstone. 1991: p. 1290-1307.

4. Hoyer LW, Breckenridge RT. Immunologic studies of antihemophilic factor (AHF, factor VIII). Blood 1968;32:962–971.

5. Stevan MM. Madhok YR, Forbes CD, Sturrock RD. Haemophilic arthritis. Quat J of Med 1986;58:181- 197.

6. Agarwal et al. Classical haemophilia. J Assoc Physicians India. 1981;29:120-130.

7. Kitchens CS. Occult haemophilia. Johns Hopkins Medical J. 1980;746:255-259.

8. Takim and Shrivastava. A current situation of regular replacement therapy for haemophilia in Japan. Hamophila 2009.15.78-82.

9. Shanthala Devi AM, Sitalakshmi S. Shrikrisha A, Damodar p, Mathew T, Ernest JP. Profile of inherited bleeding disorders in teaching hospital. Indian J hematol and blood transf 1999;17:16-18.

10. Rodgers GM and Charles. Inherited coagulation disorders. Wintrobes clinical hematology. Vol II 12th edition Philadelphia.2009p.1379-1416.



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Best Article Award

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A study by Muhas C. et al. entitled \"Study on Knowledge & Awareness About Pharmacovigilance Among Pharmacists in South India\" is awarded Best article for Vol 14 issue 22
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A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
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A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
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A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
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A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award


A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" from Vol 13 issue 06 received Emerging Researcher Award


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