A Comprehensive Guide to Biomarkers for Leukaemia (All Types)

Biomarkers are the breadcrumbs clinicians follow to diagnose, stage, treat, and monitor leukemia. Think of them as a living map of the disease. Some are simple, like a complete blood count showing too many white cells. Others are highly specific, pinpointing a single DNA change that predicts which therapy is likely to work. This guide walks through the major biomarkers used across all leukemia types, how they are interpreted in practice, and why they matter for real decisions, with science kept accurate and the language plain.

What Counts as a Biomarker in Leukemia

In leukemia, biomarkers capture measurable traits of the cancer or the body's response to it. They range from routine lab values to molecular fingerprints.¹ Blood and bone marrow analyses reveal key signals such as blast percentage, abnormal counts, and biochemical stress indicators like LDH.² Flow cytometry outlines the immunophenotype, identifying cell-surface proteins that define the leukemia's lineage and subtype.³ Cytogenetic testing detects chromosomal gains, losses, or translocations using karyotype or FISH,⁴ while molecular methods such as PCR and next-generation sequencing uncover mutations and fusion genes driving the disease.⁵

Beyond identifying the disease, biomarkers quantify residual leukemia cells after therapy (minimal or measurable residual disease, MRD)⁶ and gauge host factors like infection risk, organ stress, or coagulation abnormalities. Each biomarker serves a different purpose: diagnosis, classification, therapy guidance, or treatment tracking. Their power comes from being used together.

How Biomarkers Shape the Care Journey

Across leukemia types, biomarkers support five major decisions: establishing the diagnosis, estimating relapse risk, guiding treatment selection, tracking response, and ensuring safety. They determine whether a case is acute or chronic, myeloid or lymphoid, and help clinicians tailor therapy intensity based on risk.¹

Once treatment begins, biomarkers help confirm whether targeted therapies are hitting their intended molecular drivers and detect signs of relapse before symptoms reappear. They also alert teams to treatment complications like tumor lysis or coagulopathy. One measure alone is rarely decisive; patterns over time tell the fuller story.

First-Line Labs at Presentation

At diagnosis, testing serves two urgent goals: confirming that leukemia is present and ensuring the patient is physiologically stable. The standard first-line panel includes a complete blood count with differential and smear to quantify blasts, alongside a comprehensive metabolic panel, LDH, uric acid, phosphate, and electrolytes to assess tumor lysis risk.²

Coagulation studies (PT/INR, aPTT, fibrinogen, D-dimer) identify dangerous clotting abnormalities such as those seen in acute promyelocytic leukemia (APL).⁷ Clinicians may also order blood type, infection screens, or imaging based on symptoms. Acute leukemias are diagnosed when blasts make up roughly 20% or more of bone marrow cells.⁸ Chronic forms evolve more slowly, with elevated counts but fewer blasts.

Immunophenotyping by Flow Cytometry

Flow cytometry paints a molecular portrait of leukemia cells by mapping proteins on their surface and within the cytoplasm. It anchors diagnosis, distinguishes subtypes, and increasingly contributes to prognosis.³ In B-cell acute lymphoblastic leukemia (B-ALL), cells typically express CD19, CD22, and CD79a, often with CD10 and the nuclear marker TdT. T-cell ALL shows CD3, CD7, and CD2 expression, sometimes alongside CD1a, CD4, or CD8. Acute myeloid leukemia (AML) features markers such as CD13, CD33, CD117, and myeloperoxidase positivity, while chronic lymphocytic leukemia (CLL) reveals co-expression of CD5 and CD23 on B cells with dim CD20.⁹ Hairy cell leukemia often carries CD103 and annexin A1, which supports its characteristic appearance.

Flow cytometry also drives MRD tracking, capable of finding one leukemia cell among thousands of healthy ones in expert laboratories.⁶

Cytogenetics: Big Genomic Moves with Big Clinical Impact

Chromosome-level changes often define specific leukemia entities. Karyotyping scans the genome broadly, while FISH targets hallmark rearrangements.⁴ These findings help predict risk and direct therapy intensity. Examples include BCR-ABL1 (t(9;22)), the Philadelphia chromosome defining chronic myeloid leukemia and some ALL cases;¹⁰ PML-RARA (t(15;17)), the fusion driving APL;⁷ and RUNX1-RUNX1T1 (t(8;21)) or CBFB-MYH11 (inv(16)), which characterize favorable-risk AML.¹¹ Other recurrent changes such as KMT2A (MLL) rearrangements, hyperdiploidy or ETV6-RUNX1 in pediatric ALL,¹² and del(17p) in CLL refine prognosis.⁹

Guidelines from organizations such as the European LeukemiaNet and NCCN use these cytogenetic signatures to assign risk categories and recommend treatment strategies.¹¹ When a translocation drives the disease, it can also be used as a PCR target for MRD monitoring.

Molecular Biomarkers: Genes with Outsized Influence

Next-generation sequencing reveals dozens of genes with actionable meaning. In AML, NPM1, FLT3, CEBPA, IDH1/2, and TP53 mutations often guide decision-making.¹³ NPM1 positivity without FLT3-ITD predicts better outcomes, while FLT3-ITD confers relapse risk that can be mitigated with targeted inhibitors.¹⁴ CEBPA mutations, when biallelic, signal favorable biology, whereas TP53 mutations forecast resistance.¹³

In ALL, gene fusions like BCR-ABL1 and CRLF2- or JAK-STAT-driven lesions define distinct subtypes that respond to specific kinase inhibitors.¹² IKZF1 deletions raise relapse risk, especially if MRD persists after induction. In CLL, the mutational status of IGHV divides slow-growing from aggressive disease, and TP53, NOTCH1, and SF3B1 variants guide whether traditional chemoimmunotherapy will work.⁹ CML is driven by BCR-ABL1; quantitative PCR tracks transcript levels over time, and resistance mutations such as T315I can direct next-line therapy choices.¹⁰

Measurable Residual Disease (MRD)

MRD testing looks for what the microscope misses. Using flow cytometry, PCR, or sequencing, clinicians can detect one cancer cell among tens of thousands of normal ones. It is one of the strongest predictors of relapse across major leukemias.⁶ In ALL, MRD after induction and consolidation defines risk; 0.01% is a common threshold for positivity.¹² In AML, flow cytometry at 0.1% sensitivity can reveal residual disease, while NPM1 or fusion-gene PCR adds precision.¹⁵ CML care hinges on transcript tracking: MR3 (≤0.1%), MR4 (≤0.01%), and MR4.5 (≤0.0032%) indicate progressively deeper remissions.¹⁰ Because assay standards differ, results should be compared only across properly calibrated laboratories; for example, CML PCR results must align with the International Scale.¹⁰

Serum and Functional Biomarkers

Not all useful markers come from the genome. Routine serum tests capture how the body responds to leukemia or therapy. Elevated LDH signals rapid cell turnover, while uric acid, phosphate, and creatinine levels reflect tumor lysis and kidney stress.² Fibrinogen and D-dimer patterns can reveal coagulopathy in APL.⁷ Beta-2 microglobulin levels often mirror disease activity in CLL,⁹ and liver enzymes help track therapy safety. These contextual markers can confirm effective cytoreduction and indicate treatment tolerability.

Biomarkers by Major Leukemia Type

In AML, diagnosis hinges on blasts with myeloid features, while cytogenetic and molecular markers such as NPM1, FLT3, and CEBPA define prognosis.¹³ APL, a subtype of AML, is identified by PML-RARA and requires urgent coagulation support due to bleeding risk.⁷

Other major leukemia types include:

• Acute Lymphoblastic Leukemia (ALL): relies on lineage-specific flow cytometry and genetic subtyping;¹² Ph-positive ALL integrates BCR-ABL1 transcript tracking with MRD results to tailor therapy.
• Chronic Lymphocytic Leukemia (CLL): combines persistent lymphocytosis with characteristic immunophenotype; IGHV and TP53 mutations guide prognosis and therapy choice,⁹ and FISH results refine risk classification.
• Chronic Myeloid Leukemia (CML): defined by BCR-ABL1 and monitored through quantitative PCR milestones.¹⁰

Even rare leukemias can hinge on a single genetic clue; BRAF V600E in hairy cell leukemia¹⁶ or TCL1 involvement in T-prolymphocytic leukemia can confirm diagnosis and guide therapy.

Assay Limitations and Clinical Context

Every test has boundaries. Age-related clonal hematopoiesis can mimic disease, while low variant-allele frequencies may reflect technical noise.¹⁷ Flow MRD sensitivity depends on lab expertise and antibody panels.⁶ FISH detects known rearrangements but may miss unexpected chromosomal complexity.⁴ To ensure accuracy, results from accredited laboratories that participate in quality assessment programs are preferred.¹⁸

Life Stage and Sex-Specific Factors

Interpretation shifts with life stage. Children often have distinct, prognostically powerful genetic subtypes of ALL.¹² Older adults show more background clonal hematopoiesis, which complicates NGS data. Pregnancy introduces physiologic leukocytosis and changes in plasma volume that require individualized testing plans coordinated with maternal–fetal medicine specialists.

After Treatment Begins

Once therapy starts, biomarkers pivot from diagnosis to monitoring response and safety. Falling blast counts, LDH, and uric acid suggest successful cytoreduction, while recovering neutrophils and platelets signal marrow recovery. MRD checkpoints confirm whether remission is deepening.⁶ Organ function tests continue throughout treatment to manage toxicity and adjust dosing safely.

Questions to Ask Your Care Team

Patients benefit from understanding the markers being tracked. You might ask which cytogenetic or molecular features define your subtype, how frequently MRD will be assessed, what sensitivity the test achieves, and whether your lab participates in external quality programs.¹⁸ These questions turn biomarker data into meaningful guidance rather than numbers in isolation.

Examples of Biomarker-Driven Decisions

PML-RARA in APL triggers immediate molecular confirmation and coagulation support.⁷ NPM1 mutation status in AML informs MRD tracking.¹⁵ BCR-ABL1 trends shape therapy intensity in Ph-positive ALL.¹² In CLL, unmutated IGHV or TP53 disruption steers clinicians away from older chemotherapies.⁹ For CML, maintaining BCR-ABL1 below 0.1% at twelve months signals excellent control.¹⁰

Data Quality and Standardization

Two patients with the same diagnosis can receive different readouts if their assays differ. Reference scales, gene panel depth, and laboratory experience all influence interpretation.⁵ For high-stakes choices, such as transplant eligibility or stopping targeted therapy, confirmatory testing at an experienced center provides essential reassurance. Treatment-free remission in CML, for instance, requires sustained deep molecular response verified across multiple time points by consistently standardized methodology.¹⁹

What Biomarkers Cannot Do

Even the most sophisticated biomarker cannot guarantee an outcome. A favorable mutation does not ensure cure, and MRD negativity does not mean every malignant cell is gone. These tests quantify probability, not certainty. They are tools for decision-making within clinical context, not verdicts.

Everyday Factors That Skew Results

Common variables can distort results: infections that temporarily elevate white counts, recent transfusions that dilute marrow cells, biotin supplements that interfere with immunoassays, and transport delays that degrade RNA. Communicating recent medications, supplements, and symptoms helps ensure accurate interpretation.

The Bottom Line

Biomarkers translate the complexity of leukemia into measurable signals that guide care from diagnosis through remission. They define subtype, clarify prognosis, and monitor response at levels below what the eye can see. The most meaningful insights come not from any single value but from trends over time, interpreted alongside a knowledgeable care team and validated laboratory data. That is how modern leukemia care turns molecular detail into durable outcomes.