Minimally Invasive Method of Measuring Anti-Müllerian Hormone Level in Dried Blood Spots of Indonesian Infertile Population

Sini, Ivan and Handayani, Nining and Tjahjadi, Ricky and Wulandari, Tri Aprilliana and Afadlal, Szeifoul and Polim, Arie Adrianus and Sirait, Batara I. and Suheimi, Irham and Boediono, Arief (2024) Minimally Invasive Method of Measuring Anti-Müllerian Hormone Level in Dried Blood Spots of Indonesian Infertile Population. FERTILITY & REPRODUCTION : The Official Journal of the Asia Pacific Initiative on Reproduction (ASPIRE), 6 (1). pp. 42-48. ISSN 2661-3174

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Abstract

Penelitian ini bertujuan untuk mengevaluasi kadar hormon anti-Müllerian (AMH) menggunakan sampel darah kering (dried blood spots, DBS) dibandingkan dengan serum darah. Sampel DBS dan serum dikoleksi dari 109 wanita infertil yang akan menjalani program bayi tabung. Kadar AMH sampel DBS dianalisis menggunakan reagen kit ELISA DBS AMH (Ansh Lab) sedangkan kadar AMH sampel serum dianalisis menggunakan Elecsys® AMH plush Assay (Roche). Rerata kadar AMH pada sampel DBS adalah 4,02 ± 3,89 ng/mL (0,01 – 17,3 ng/mL), sedangkan rerata kadar AMH pada sampel serum adalah 2,44 ± 2,29 ng/mL (0,01–10,8 ng/ mL). Diperoleh akurasi sebanding antara kedua sampel dalam mengukur AMH kadar rendah dibandingkan terhadap jumlah folikel basal sebagai pembanding klinis (AUC 86%, titik potong ≤1.95 ng/mL, R = 0.67, P < 0.001 vs. 85%, titik potong ≤1.15 ng/mL, R = 0.66, P < 0.001, masing-masing untuk sampel DBS dan serum). Sampel DBS juga mencapai kesesuaian yang sangat baik dengan sampel serum untuk menentukan AMH kadar rendah (AUC 99% vs. 93%). Regresi linear Spearman yang kuat (R = 0.92, p < 0.001) dan tingkat kesesuaian yang cukup (94.5%, perbedaan rerata −1.6 (2.7 sampai −5.9)) diamati pada kedua group sampel dengan rumus matematika regresi Passing dan Bablok y = −0.12+1.80×. Selang kepercayaan untuk nilai intercept ((−0.12, 95% CI [−0.29 to −0.01]) dan slope (1.80, 95% CI [1.66–1.94]) menunjukan adanya perbedaan konstan dan proporsional antara kedua sampel, namun tidak bermakna (P = 0.893). Dapat disimpulkan bahwa pengukuran kadar AMH dengan sampel DBS dapat menjadi pilihan untuk evaluasi cadangan folikel ovarium melalui koleksi sampel jarak jauh yang lebih nyaman. Kata kunci: Hormon Anti-Mullerian, Sampel darah kering, Fertilisasi in-vitro. / This study aims to evaluate the certainty of anti-Müllerian hormone (AMH) levels in dried blood spots (DBS) compared to serum AMH. A total of 109 infertile women who planned to undergo their controlled ovarian stimulation cycle were recruited. Serum and DBS samples were collected from each patient for the quantification of AMH levels. Serum AMH was measured with an automated clinical chemistry assay (Roche) while DBS AMH was measured using the Ansh Labs DBS AMH ELISA reagent kit. Mean AMH level in DBS was 4.02 ± 3.89 ng/mL, which ranged from 0.01 to 17.3 ng/mL, while the mean AMH in serum samples was 2.44 ± 2.29 ng/mL (0.01–10.8 ng/mL). Comparable accuracy for low AMH quantification was observed between the two methods over basal antral follicle counts (AFC) as a clinical comparator (AUC 86%, cutoff ≤1.95 ng/mL, R = 0.67, P < 0.001 vs. 85%, cutoff ≤1.15 ng/ mL, R = 0.66, P < 0.001, respectively for DBS and serum sample). Likewise, DBS AMH achieved high concordance for low AMH discrimination in comparison to serum AMH method (AUC 99% vs. 93%, respectively). A strong linear Spearman regression (R = 0.92, P < 0.001) and sufficient agreement (94.5%, mean difference of −1.6, 2.7 to −5.9) was demonstrated among studied groups with mathematical Passing and Bablok regression equation of y = −0.12+1.80×. The confidence interval of intercept (−0.12, 95% CI [−0.29 to −0.01]) and slope (1.80, 95% CI [1.66–1.94]) indicated a constant and proportional difference between the two methods but it was not significant (P = 0.893). AMH measurement in DBS could be useful as an alternative approach to screening ovarian reserve through a more convenient remote sample collection. Keywords: Anti-Müllerian Hormone; Dried Blood Spot; In-vitro Fertilization.

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