Haemoglobin Electrophoresis Test Explained: Normal Range, HbA2, HbF, Thalassemia & Sickle Cell (India 2026) | हीमोग्लोबिन इलेक्ट्रोफोरेसिस टेस्ट गाइड
Haemoglobin Electrophoresis & HPLC Test Explained: Thalassaemia, Sickle Cell & Report Reading (India 2026)
हीमोग्लोबिन इलेक्ट्रोफोरेसिस और HPLC टेस्ट: थैलेसीमिया, सिकल सेल और रिपोर्ट पढ़ने की पूरी गाइड
You have been told your CBC (Complete Blood Count) shows mild anaemia that does not respond to iron — or perhaps your haemoglobin is persistently low and your MCV is small, yet your iron stores and ferritin are normal. In these situations, your doctor orders a Haemoglobin Electrophoresis or HPLC (High-Performance Liquid Chromatography) test to identify exactly which types of haemoglobin your red blood cells contain. This test is the definitive investigation for diagnosing thalassaemia (both trait and disease), sickle cell anaemia, sickle cell trait, and other haemoglobin variants that are extremely prevalent in India.
India carries one of the world's largest burdens of haemoglobinopathies — an estimated 40 million thalassaemia carriers (beta thalassaemia trait) and approximately 14 million sickle cell trait carriers, with approximately 10,000–12,000 thalassaemia major babies born every year. Understanding your haemoglobin electrophoresis report — including what terms like HbA2 elevated, HbS present, or HbF elevated actually mean — is critical for patients, families, and especially couples planning pregnancy. For reading lab reports in general, see our beginner's guide to blood test reports.
भारत में अनुमानित 4 करोड़ थैलेसीमिया वाहक (बीटा थैलेसीमिया ट्रेट) और 1.4 करोड़ सिकल सेल ट्रेट वाहक हैं। हीमोग्लोबिन इलेक्ट्रोफोरेसिस / HPLC टेस्ट उनकी पहचान करने का निश्चित तरीका है। यह गाइड सरल अंग्रेजी और हिंदी में इसे समझाती है। Table of Contents / विषय सूची
- What Is Haemoglobin Electrophoresis? / टेस्ट क्या है?
- Normal Haemoglobin Types & Reference Values
- Thalassaemia — Trait vs Disease / थैलेसीमिया
- Sickle Cell Anaemia & Sickle Cell Trait / सिकल सेल
- Other Haemoglobin Variants in India
- When Is This Test Ordered?
- HPLC vs Standard Electrophoresis
- Frequently Asked Questions / FAQ
What Is Haemoglobin Electrophoresis? / हीमोग्लोबिन इलेक्ट्रोफोरेसिस क्या है?
Haemoglobin is the iron-containing protein in red blood cells that carries oxygen. In healthy adults, haemoglobin exists in several distinct molecular forms — each with slightly different amino acid sequences in their protein chains. The most important forms are HbA (the dominant adult form), HbA2 (a minor adult form), and HbF (foetal haemoglobin, normally present only in small amounts in adults). In genetic haemoglobin disorders like thalassaemia and sickle cell disease, the proportions of these forms change — or abnormal variants like HbS or HbE appear. Haemoglobin electrophoresis and HPLC are laboratory techniques that separate and quantify these different haemoglobin types from a single blood sample.
हीमोग्लोबिन लाल रक्त कोशिकाओं में ऑक्सीजन वाहक प्रोटीन है। स्वस्थ वयस्कों में यह मुख्यतः HbA, HbA2 और HbF के रूप में होता है। थैलेसीमिया और सिकल सेल जैसे आनुवंशिक विकारों में इनकी मात्रा बदल जाती है या असामान्य वेरिएंट जैसे HbS या HbE प्रकट होते हैं। HPLC और इलेक्ट्रोफोरेसिस इन्हें अलग कर उनकी मात्रा मापते हैं।Normal Haemoglobin Types & Reference Values / सामान्य हीमोग्लोबिन प्रकार
*Reference ranges may vary slightly between HPLC platforms (Bio-Rad Variant II, Tosoh G8) and standard alkaline/acid electrophoresis. Always check the reference range on your specific report. Results must be interpreted by a haematologist in the context of the CBC and clinical picture.
*HPLC प्लेटफॉर्म के बीच थोड़ी भिन्नता हो सकती है। अपनी रिपोर्ट की सीमा और अपने हेमेटोलॉजिस्ट से परामर्श करें।| Haemoglobin Type | Normal % in Adults | What it is / Clinical significance |
|---|---|---|
| HbA सामान्य वयस्क हीमोग्लोबिन |
95 – 98% | The dominant adult haemoglobin. Made of two alpha and two beta chains. Reduced in beta thalassaemia; absent in beta thalassaemia major. |
| HbA2 माइनर एडल्ट हीमोग्लोबिन |
1.5 – 3.5% | Minor adult form. The most important marker on the report. Above 3.5% = beta thalassaemia trait. Above 5–7% = possible delta-beta thalassaemia or compound heterozygote. |
| HbF फोएटल हीमोग्लोबिन |
< 1% (adults) | Foetal haemoglobin, normally almost absent in adults. Elevated HbF (above 2–5%) seen in beta thalassaemia trait, sickle cell disease (protective), hereditary persistence of foetal haemoglobin (HPFH), and delta-beta thalassaemia. |
| HbS सिकल सेल हीमोग्लोबिन |
Absent in normal adults | Abnormal haemoglobin caused by a point mutation (glutamic acid → valine at position 6 of the beta chain). Present in sickle cell trait (HbAS ~35–40%) and sickle cell disease (HbSS ~80–95%). |
| HbE HbE वेरिएंट |
Absent in normal adults | Common in eastern India, Northeast, West Bengal, Odisha. HbE trait is benign. HbE disease (HbEE) causes mild anaemia. HbE/beta thalassaemia compound is clinically significant and can be as severe as thalassaemia major. |
| HbC, HbD, HbG | Absent in normal adults | Less common variants. HbD (Punjab) seen in Punjab and western India. These are identified on electrophoresis and distinguished by their migration pattern and HPLC retention time. |
Thalassaemia — Trait vs Disease / थैलेसीमिया
Thalassaemia is a group of inherited blood disorders in which the body makes an abnormal form or insufficient amounts of haemoglobin, leading to anaemia of varying severity. India has two main types: beta thalassaemia (deficiency or absence of beta globin chains — the most common form in India) and alpha thalassaemia (deficiency of alpha chains). The distinction between being a carrier (thalassaemia trait) and having the disease (thalassaemia major or intermedia) is critical — and is what haemoglobin electrophoresis definitively establishes.
थैलेसीमिया आनुवंशिक रक्त विकारों का एक समूह है जिसमें शरीर असामान्य या अपर्याप्त हीमोग्लोबिन बनाता है। भारत में मुख्यतः बीटा थैलेसीमिया आम है। वाहक (ट्रेट) और रोग (मेजर) के बीच का अंतर हीमोग्लोबिन इलेक्ट्रोफोरेसिस द्वारा निश्चित रूप से स्थापित किया जाता है।The most common haemoglobin electrophoresis finding in India. One abnormal beta globin gene inherited (heterozygous). Report pattern: HbA2 elevated (3.5–7%), HbF mildly elevated or normal, HbA normal to mildly reduced. CBC shows mild microcytic anaemia (Hb 10–12 g/dL), low MCV, low MCH, normal ferritin. Carriers are clinically well — they do not need treatment. The critical implication is for family planning: if both partners carry beta thalassaemia trait, each pregnancy has a 25% chance of thalassaemia major.
Both beta globin genes are abnormal (homozygous or compound heterozygous). Report pattern: HbA very low or absent, HbF markedly elevated (40–90%), HbA2 variable. Severe anaemia requiring lifelong monthly blood transfusions. Without treatment, leads to serious complications (iron overload, organ damage). Preventable through carrier screening before pregnancy and prenatal diagnosis. Bone marrow transplantation can be curative in eligible patients.
Alpha globin gene deletions cause a range from silent carrier (one gene deleted — completely normal) to Hb Bart hydrops fetalis (four genes deleted — incompatible with life). Alpha thalassaemia trait (two genes deleted) shows low MCV and mild anaemia but a normal HbA2 on electrophoresis — this is why standard electrophoresis can miss it. Alpha thalassaemia diagnosis requires DNA analysis (alpha globin gene mutation testing). Suspected when CBC shows low MCV/MCH with normal HbA2 and normal iron studies.
In West Bengal, Odisha, Assam, and northeast India, the HbE variant is common. HbE trait (one HbE gene) causes mild microcytic anaemia but is otherwise benign. The combination of HbE trait + beta thalassaemia trait (both parents carrying different variants) creates a compound heterozygote HbE/beta thalassaemia — which can be clinically as severe as thalassaemia major, requiring regular transfusions. Eastern Indian couples undergoing pre-pregnancy screening must specifically test for HbE and beta thalassaemia trait.
Sickle Cell Anaemia & Sickle Cell Trait / सिकल सेल
Sickle cell disease is caused by a point mutation in the beta globin gene that changes the amino acid glutamic acid to valine at position 6, producing an abnormal haemoglobin called HbS. When HbS molecules are deoxygenated, they polymerise and distort the red blood cell into a rigid crescent (sickle) shape — causing vascular occlusion, pain crises, haemolytic anaemia, and progressive organ damage. Sickle cell disease is particularly prevalent in central and peninsular India — Madhya Pradesh, Chhattisgarh, Odisha, Maharashtra, and tribal communities across India carry some of the highest sickle cell gene frequencies in the world.
सिकल सेल रोग बीटा ग्लोबिन जीन में एक बिंदु उत्परिवर्तन के कारण होता है। मध्य प्रदेश, छत्तीसगढ़, ओडिशा, महाराष्ट्र और आदिवासी समुदायों में यह विशेष रूप से प्रचलित है।Report: HbA approximately 55–60%, HbS approximately 35–40%, HbA2 normal, HbF normal. Carriers have one normal beta gene and one HbS gene. They are generally clinically well — no pain crises, normal lifespan. May have mild symptoms at extreme altitude, severe dehydration, or during general anaesthesia (rare). Critical for family planning: if both partners carry sickle cell trait, each pregnancy has a 25% chance of sickle cell disease. The National Sickle Cell Anaemia Elimination Mission (2023) mandates screening in high-prevalence states.
Report: HbS dominant (80–95%), HbA absent, HbF variable (5–20%), HbA2 normal. Both beta globin genes carry the HbS mutation (homozygous). Clinical features: severe haemolytic anaemia, episodic acute pain crises (vaso-occlusive crises), acute chest syndrome, splenic sequestration, stroke, avascular necrosis, chronic organ damage. Treatment: hydroxyurea (reduces HbF and crisis frequency), regular transfusions, supportive care. Curative options include bone marrow transplantation and gene therapy (now available in India at select centres).
Other Haemoglobin Variants & Report Patterns
A benign condition where HbF persists in unusually high amounts in adults (above 10–25%) without clinical symptoms. Report shows elevated HbF with corresponding reduction in HbA, but normal or near-normal HbA2. When HPFH coexists with sickle cell gene, the high HbF actually ameliorates the clinical severity of sickle cell disease — one reason why HPFH is clinically important to identify correctly.
HbD (beta chain mutation) is found predominantly in Punjab and northwest India. HbD trait is clinically benign. HbD/beta thalassaemia compound causes moderate to severe anaemia. HbD co-migrates with HbS on alkaline electrophoresis — which is why HPLC or acid electrophoresis is important to distinguish the two, as HbS and HbD require very different management.
A deletion that removes both the delta and beta globin genes. Report: HbF markedly elevated (5–20% in trait, much higher in homozygous disease), HbA2 normal or low (unlike beta thalassaemia trait where HbA2 is high — this is a key distinguishing feature). Delta-beta thalassaemia trait is clinically benign but important to identify for genetic counselling.
HbC is rare in India (more common in West Africa) but seen occasionally in travellers and immigrants. HbC disease causes mild haemolytic anaemia with target cells on CBC peripheral smear. HbG Philadelphia is an alpha chain variant. All these are identified by their characteristic retention times on HPLC and migration positions on electrophoresis. A haematologist review is recommended for any unexpected variant band on the report.
When Is This Test Ordered in India? / कब यह टेस्ट करवाना चाहिए?
Haemoglobin electrophoresis is ordered in specific clinical scenarios where a CBC and basic iron studies are insufficient. Understanding when the test is appropriate — and why — helps patients engage more meaningfully with their diagnosis.
हीमोग्लोबिन इलेक्ट्रोफोरेसिस विशिष्ट नैदानिक स्थितियों में मंगाया जाता है जहां CBC और बुनियादी आयरन अध्ययन पर्याप्त नहीं होते।The most common indication in India. When a CBC shows low Hb with low MCV and MCH, but the anaemia does not improve after 2–3 months of iron supplementation — and iron studies (serum ferritin, TIBC) are normal or near-normal — beta thalassaemia trait is the most likely explanation. Electrophoresis confirms the diagnosis and prevents unnecessary prolonged iron supplementation.
Any couple planning pregnancy should ideally have haemoglobin electrophoresis done — particularly if either partner has a family history of anaemia, thalassaemia, sickle cell disease, or comes from a high-prevalence community (central India, eastern India, coastal states, tribal communities). If one partner is found to be a carrier, the other must be tested immediately. This prevents the birth of a child with thalassaemia major or sickle cell disease. Prenatal diagnosis by chorionic villus sampling (CVS) at 10–13 weeks or amniocentesis at 15–18 weeks is available if both partners carry the same mutation.
All pregnant women with anaemia should have haemoglobin electrophoresis as part of the antenatal workup — alongside thyroid function, Vitamin D, and the beta hCG. The National Health Mission recommends testing all pregnant women in high-prevalence areas. Ideally, screening should happen before pregnancy — but first-trimester identification still allows prenatal diagnosis in time for informed decision-making. The second-trimester triple marker screen does not detect haemoglobin disorders — a separate electrophoresis is needed.
Any patient with persistently low MCV and MCH on CBC, normal serum ferritin, and no response to iron — even without overt anaemia — should have haemoglobin electrophoresis. Similarly, anyone with a family history of thalassaemia, sickle cell disease, repeated blood transfusions in a sibling, or unexplained haemolytic anaemia in a first-degree relative should be screened. The test is also done when a blood transfusion is being planned, to match compatible haemoglobin types.
HPLC vs Standard Electrophoresis — Which Is Better?
| Feature | Standard Haemoglobin Electrophoresis | HPLC (High-Performance Liquid Chromatography) |
|---|---|---|
| Principle | Separation of Hb types by electrical charge on cellulose acetate membrane | Separation by chemical interaction (ion-exchange chromatography) with quantification by UV absorbance |
| Sensitivity / accuracy | Good for HbS, HbC, HbA2 estimation; may miss some variants | Higher sensitivity — better at detecting subtle HbA2 elevation, rare variants, HbE |
| HbA2 quantification | Estimated — less precise | Precisely quantified — more reliable for borderline HbA2 (3.5–4.0%) |
| HbD vs HbS distinction | Both migrate at same position on alkaline electrophoresis — cannot be distinguished without acid electrophoresis | Distinguished by different retention times — single run |
| Speed | 3–6 hours | Less than 1 hour — automated |
| Availability in India | Widely available including district labs | Available at major cities and NABL-accredited labs; government thalassaemia centres |
| Current recommendation | Acceptable for initial screening in resource-limited settings | Preferred method at all tertiary centres per Indian Academy of Paediatrics and haematology guidelines |
✅ Book Haemoglobin Electrophoresis / HPLC Test — Home Collection Available
If your CBC shows microcytic anaemia with normal iron studies, or your doctor has recommended thalassaemia screening before pregnancy, this is the definitive next test:
Affiliate link: I may earn a small commission at no extra cost to you. Prices as of April 2026. Government thalassaemia centres and district hospitals offer this test at low or no cost. If you are pregnant or planning pregnancy, access government facilities first for free testing under NHM.
हीमोग्लोबिन इलेक्ट्रोफोरेसिस — थैलेसीमिया ट्रेट और सिकल सेल वाहक की पहचान के लिए। सरकारी थैलेसीमिया केंद्रों और जिला अस्पतालों में यह टेस्ट मुफ्त या बहुत कम लागत पर उपलब्ध है। Nutritional Support for Blood Health — Recommended Supplements
MuscleBlaze MB-Vite Daily Multivitamin — 51 Ingredients (60 Tablets)
Comprehensive daily multivitamin for overall health support — useful for general nutritional maintenance in thalassaemia trait carriers and those with mild anaemia. Includes iron, B-vitamins, zinc, and antioxidants. Always consult your haematologist before starting any supplement — high-dose iron supplementation is contraindicated in thalassaemia; the correct supplement type and dose depends on your specific haemoglobin pattern and iron stores.
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INLIFE Vitamin B12 (Methylcobalamin) 1500 mcg with ALA & Folic Acid
Folic acid is routinely prescribed by haematologists for thalassaemia trait carriers and patients with haemolytic anaemias — because rapid red cell turnover increases folate demand. Folic acid supports red blood cell production and reduces haemolytic anaemia severity. Combined with Vitamin B12 to prevent masked B12 deficiency. Always consult your doctor before starting any supplement — thalassaemia major patients have specific nutritional requirements managed under specialist supervision.
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Related Tests / संबंधित जांचें
These tests are commonly ordered alongside or after Haemoglobin Electrophoresis in India:
भारत में हीमोग्लोबिन इलेक्ट्रोफोरेसिस के साथ या बाद में ये जांचें अक्सर करवाई जाती हैं:Frequently Asked Questions / अक्सर पूछे जाने वाले सवाल
HbA2 is a minor adult haemoglobin variant — normally it constitutes 1.5–3.5% of total haemoglobin. An HbA2 above 3.5% (especially 3.5–7%) on an electrophoresis or HPLC report is the classic diagnostic marker of beta thalassaemia trait (beta thalassaemia carrier status). This means you have inherited one abnormal beta globin gene (from one parent) and one normal gene. As a carrier, you are clinically well — you do not have thalassaemia disease, you do not need treatment, and you can live a completely normal life. The critical implication is for family planning: if your partner also has beta thalassaemia trait, each pregnancy carries a 25% risk of producing a child with thalassaemia major. If HbA2 is above 7%, further evaluation for compound heterozygosity or HbE/beta thalassaemia may be needed.
उत्तर: HbA2 3.5% से ऊपर बीटा थैलेसीमिया ट्रेट (वाहक) का क्लासिक संकेत है। वाहक नैदानिक रूप से स्वस्थ होते हैं और उन्हें उपचार की जरूरत नहीं। महत्वपूर्ण निहितार्थ गर्भावस्था योजना के लिए है।No — a thalassaemia trait (carrier) diagnosis is not a reason to worry about your own health. Carriers are clinically well throughout their lives. They may have mild microcytic anaemia that does not respond to iron — which is simply a feature of the trait, not a disease requiring treatment. You do not need iron supplements for this mild anaemia (adding iron to a thalassaemia carrier who is already iron-replete can actually cause iron overload). The one critical action is to inform your partner: if you are in a relationship and planning to have children, your partner must also be tested. If both partners are beta thalassaemia carriers, genetic counselling before conception is essential. Prenatal diagnosis (by chorionic villus sampling or amniocentesis) can determine the child's status in early pregnancy, allowing informed decision-making.
उत्तर: नहीं — थैलेसीमिया ट्रेट (वाहक) निदान अपने स्वास्थ्य के बारे में चिंता का कारण नहीं है। वाहक जीवन भर नैदानिक रूप से स्वस्थ रहते हैं। सबसे महत्वपूर्ण कदम: अपने साथी को सूचित करें और दोनों की जांच करवाएं।No — fasting is not required for haemoglobin electrophoresis or HPLC. Haemoglobin variants and their proportions are not affected by food intake. The test can be done at any time of day after eating normally. However, there is one important consideration: the test should ideally not be done within 3 months of a blood transfusion, as transfused donor blood will contain the donor's haemoglobin types — mixing these with the patient's own haemoglobin can make the results difficult to interpret or falsely normalise an abnormal pattern. Inform your doctor and lab if you have received a recent transfusion.
उत्तर: नहीं — हीमोग्लोबिन इलेक्ट्रोफोरेसिस के लिए उपवास आवश्यक नहीं। महत्वपूर्ण: रक्त आधान (ब्लड ट्रांसफ्यूजन) के 3 महीने के भीतर परीक्षण से बचें — दाता हीमोग्लोबिन परिणामों को प्रभावित कर सकता है।Thalassaemia trait (carrier) means you have inherited one abnormal beta globin gene and one normal gene (heterozygous). You produce enough normal haemoglobin to be clinically well — only mild, non-progressive anaemia. No treatment needed. Thalassaemia major means you have inherited two abnormal beta globin genes (homozygous or compound heterozygous) — one from each parent. Both copies of the gene are defective, so the body cannot produce adequate normal haemoglobin. This results in severe, life-threatening anaemia from early childhood requiring monthly blood transfusions, chelation therapy to remove excess iron from transfused blood, and ideally a bone marrow transplant. Thalassaemia major is entirely preventable by screening both parents for carrier status before conception — if both are carriers, prenatal diagnosis can identify whether the foetus has thalassaemia major.
उत्तर: थैलेसीमिया ट्रेट = एक असामान्य जीन (नैदानिक रूप से स्वस्थ, कोई उपचार नहीं)। थैलेसीमिया मेजर = दो असामान्य जीन (गंभीर एनीमिया, जीवन भर मासिक रक्त आधान)। मेजर पूरी तरह से रोकथाम योग्य है।A normal haemoglobin electrophoresis result (normal HbA, HbA2, HbF, no abnormal variants) combined with a persistently low MCV and low MCH on CBC — and normal iron studies — is the classic presentation of alpha thalassaemia trait. Alpha thalassaemia is caused by deletions of alpha globin genes, and these deletions do not produce any abnormal haemoglobin band or change in HbA2 — which is why standard electrophoresis and HPLC cannot detect it. The diagnosis requires DNA analysis of the alpha globin genes (PCR-based mutation testing). Alpha thalassaemia trait is extremely common in India and is usually benign, but is important to identify for pre-pregnancy genetic counselling — particularly in couples where one or both partners come from communities with high alpha thalassaemia prevalence (South India, coastal states, tribal communities).
उत्तर: सामान्य इलेक्ट्रोफोरेसिस + लगातार कम MCV/MCH + सामान्य आयरन = अल्फा थैलेसीमिया ट्रेट की संभावना। इसका निदान DNA विश्लेषण (अल्फा ग्लोबिन जीन PCR) से होता है — इलेक्ट्रोफोरेसिस से नहीं।For thalassaemia trait carriers — no cure is needed, because they are well. For thalassaemia major patients, the traditional standard of care is lifelong regular blood transfusions (every 3–4 weeks) combined with iron chelation therapy to manage transfusional iron overload — this keeps patients well but is not curative. Bone marrow transplantation (BMT) from a matched sibling donor is the only established curative treatment — with cure rates above 90% in younger patients done at experienced centres. In India, several government and private centres perform BMT for thalassaemia, and government schemes provide partial cost coverage. Gene therapy for thalassaemia has now received approval in some countries (Zynteglo in Europe/USA) and clinical programmes are being established in India. The most effective approach at the population level remains prevention through carrier screening — testing both partners before pregnancy prevents thalassaemia major births entirely.
उत्तर: थैलेसीमिया मेजर के लिए: बोन मैरो ट्रांसप्लांट (BMT) एकमात्र स्थापित इलाज है, 90%+ सफलता। भारत में कई केंद्र BMT करते हैं। जीन थेरेपी उभर रही है। जनसंख्या स्तर पर: गर्भावस्था से पहले वाहक स्क्रीनिंग से थैलेसीमिया मेजर जन्म को पूरी तरह रोका जा सकता है।- MedlinePlus (NIH): Haemoglobin Electrophoresis — Patient Information
- Thalassaemia International Federation: TIF — Global Thalassaemia Resources
- National Health Mission, India: Sickle Cell Anaemia Elimination Mission — Government of India thalassaemia and sickle cell screening programmes.
⚠️ Medical Disclaimer / चिकित्सा अस्वीकरण
This article is for educational purposes only. Haemoglobin electrophoresis and HPLC results must always be interpreted by a qualified haematologist in the context of the CBC, iron studies, clinical history, and family background. Do not make family planning decisions or start or stop any treatment based on this guide alone. If you have received an abnormal haemoglobin electrophoresis result, consult a haematologist or genetic counsellor promptly.
यह लेख केवल शैक्षिक उद्देश्यों के लिए है। हीमोग्लोबिन इलेक्ट्रोफोरेसिस परिणाम हमेशा एक योग्य हेमेटोलॉजिस्ट द्वारा CBC, आयरन अध्ययन और नैदानिक इतिहास के संदर्भ में व्याख्या किए जाने चाहिए। असामान्य परिणाम पर तुरंत हेमेटोलॉजिस्ट से परामर्श लें।
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