1.Introduction
Polycystic ovary syndrome (PCOS) is one of the most prevalent endocrine disorders affecting women of reproductive age, with a prevalence of up to 13% in this population [1]. This multifaceted condition is characterized by a spectrum of clinical manifestations, including menstrual irregularities, hyperandrogenism (clinical or biochemical), and polycystic ovarian morphology. Despite significant advancements in understanding PCOS pathophysiology, its diagnosis remains a complex challenge requiring meticulous evaluation of clinical, biochemical, and ultrasonographic data. Updated guidelines and emerging diagnostic markers, such as anti-Müllerian hormone (AMH), continue to refine diagnostic approaches, enabling more accurate and timely identification of the syndrome.

2.Evolution of PCOS Diagnostic
CriteriaHistorically, PCOS diagnosis was guided by the Rotterdam criteria (2003), which required two of three features: oligo- or anovulation, clinical or biochemical hyperandrogenism, and polycystic ovarian morphology on ultrasound. However, with evolving scientific knowledge and clinical experience, international guidelines have undergone revisions. In 2023, updated evidence-based international guidelines for the assessment and management of PCOS were published, transitioning from consensus-based to evidence-based criteria, significantly improving diagnostic precision and standardizing patient management approaches [2]. These guidelines, still relevant in 2025, emphasize the importance of an interdisciplinary approach and consideration of individual patient characteristics.

3. Key Hormonal Markers in PCOS Diagnosis
3.1. Anti-Müllerian Hormone (AMH)Anti-Müllerian hormone (AMH), a key marker of ovarian reserve, is produced by granulosa cells of preantral and small antral follicles. In PCOS, AMH levels are significantly elevated, reflecting an increased number of small ovarian follicles [3]. The 2023 guidelines, supported by research from 2024–2025, endorse AMH as an alternative to ultrasound for diagnosing polycystic ovarian morphology in adult women [1, 4]. However, standardized AMH thresholds for PCOS diagnosis remain under discussion. Studies propose cut-off values ranging from 3.2 ng/mL to 5.055 ng/mL, noting that AMH levels may vary depending on age, body mass index, and laboratory assay methods [5, 6, 7]. Importantly, elevated AMH alone is insufficient for a PCOS diagnosis without other supporting criteria [4]. In adolescents, AMH is not recommended for PCOS diagnosis due to physiologically elevated levels during this period [8].3.2. Luteinizing Hormone (LH) and Follicle-Stimulating Hormone (FSH)Dysregulated gonadotropin secretion, particularly an elevated LH/FSH ratio, has long been considered a hallmark of PCOS. Increased pulsatile gonadotropin-releasing hormone (GnRH) activity in PCOS often results in predominant LH secretion over FSH [9]. Mean LH levels and the LH/FSH ratio are significantly higher in women with PCOS compared to healthy controls [10]. However, an elevated LH/FSH ratio is not a universal diagnostic criterion, as it is not consistently observed, particularly in women with normal body weight [11]. Its diagnostic utility is limited by high variability and dependence on menstrual cycle phase.3.3. Androgens (Testosterone, DHEA-S, Androstenedione)Hyperandrogenism is a cornerstone of PCOS, manifesting clinically (hirsutism, acne, androgenic alopecia) or biochemically (elevated blood androgen levels). Measurement of total and free testosterone is recommended for assessing biochemical hyperandrogenism [2]. Free testosterone is generally a more sensitive marker, reflecting the biologically active hormone fraction. Levels of dehydroepiandrosterone sulfate (DHEA-S) and androstenedione may also be elevated but are primarily measured to rule out other causes of hyperandrogenism, such as adrenal or ovarian tumors.

3.4. Insulin and Glucose
Insulin resistance is a key pathophysiological feature of PCOS, contributing to metabolic disturbances and hyperandrogenism [2]. However, routine measurement of insulin levels or insulin resistance indices (e.g., HOMA-IR) is not recommended for PCOS diagnosis due to their lack of specificity and absence of standardized thresholds [2]. Nonetheless, assessing metabolic parameters, such as fasting glucose and oral glucose tolerance testing, is critical for identifying carbohydrate metabolism disorders and evaluating the risk of type 2 diabetes in women with PCOS.

4. Comprehensive Approach to Diagnosis and Interpretation of Laboratory Data
Diagnosing PCOS requires a comprehensive approach, integrating detailed medical history, physical examination, clinical symptom evaluation, and interpretation of laboratory and imaging data. Given the variability in hormonal profiles and the lack of universal thresholds for certain markers like AMH, accurate interpretation of laboratory results is critical. This is particularly relevant amid evolving guidelines and the emergence of new biomarkers.In this context, services providing precise interpretation of laboratory results are invaluable for clinicians and patients. Aima Diagnostics, an innovative platform leveraging advanced artificial intelligence algorithms, facilitates the analysis and interpretation of laboratory data. It accounts for individual patient characteristics, aligns results with the latest scientific literature and clinical guidelines, and detects subtle deviations that may be overlooked in standard practice. The use of platforms like Aima Diagnostics enhances PCOS diagnostic accuracy, optimizes treatment strategies, and improves patient outcomes by providing clinicians with deeper insights into the complex hormonal landscape of the syndrome.

5. Conclusion
In 2025, PCOS diagnosis is grounded in evidence-based international guidelines that continue to evolve, incorporating novel biomarkers such as AMH. Understanding the roles of key hormonal markers—AMH, LH/FSH, androgens, and metabolic parameters—is fundamental to accurate diagnosis. A comprehensive approach, supported by advanced tools like Aima Diagnostics for laboratory data interpretation, opens new avenues for personalized medicine and significantly enhances the quality of care for women with PCOS.


6. References
[1] AAFP. (2024). Polycystic Ovary Syndrome: Assessment and Management Guidelines. American Family Physician, 110(5): 547-548.

[2] ASRM. (2023). Recommendations from the 2023 International Evidence-based Guideline for the Assessment and Management of Polycystic Ovary Syndrome. Practice Guidance.

[3] Dileep, A. (2025). Evaluating serum anti-Müllerian hormone as a diagnostic marker for polycystic ovary syndrome. PubMed.

[4] AAFP. (2024). New Diagnostic Option for Polycystic Ovarian Syndrome. AAFP Community Blog.

[5] Piltonen, T. T. (2025). Prospective validation of anti-Müllerian hormone cutoff to determine polycystic ovarian morphology. ScienceDirect.

[6] Ho, N. T. (2025). Characteristics of serum anti-Müllerian hormone levels in women with polycystic ovary syndrome. Human Reproduction, 40(Supplement_1): deaf097.129.

[7] Vale-Fernandes, E. (2025). Should anti-Müllerian hormone be a diagnosis criterion for polycystic ovary syndrome? PubMed.

[8] Peña, A. S. (2025). International evidence-based recommendations for polycystic ovary syndrome. BMC Medicine.

[9] Su, P. (2025). Physiopathology of polycystic ovary syndrome in adolescents. Ovarian Research.

[10] Akhter, S. (2025). Evaluation of Luteinizing Hormone, Follicle Stimulating Hormone and LH/FSH Ratio in Polycystic Ovary Syndrome. Bangladesh Journal of Medical Science.

[11] Pratama, G. (2024). Mechanism of elevated LH/FSH ratio in lean PCOS revisited. Nature.