HBA® Assay

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HBA® Assay 1

A diagnostic tool with dual Hyaluronan coated chambers for sperm sample evaluation

The HBA™ Assay is a diagnostic tool with dual hyaluronan coated chambers for sperm sample analysis. It allows the ability to distinguish between mature spermatozoa expressing hyaluronan receptors on their plasma membrane, and immature ones lacking those receptors.

The presence of receptors and the ability of spermatozoa to bind to hyaluronan is correlated with sperm maturity, increased chromatin integrity, decreased chances of aneuploidy, lower sperm DNA fragmentation and better sperm DNA packaging.1,2,3

The proportion of motile spermatozoa capable to bind to hyaluronan is called the Hyaluronan Binding index or HBA index. There is a correlation between the HBA index and ICSI treatment outcomes.4,5,6

Assessing the proportion of mature spermatozoa with hyaluronan receptors can be used to determine the best treatment options for the patients.

Key Features

  • Easy to implement, simple to perform
  • Sperm sample evaluation takes minutes
  • More data on sperm quality for the informed choice of treatment tactics


This product is available for sale in Europe, the U.S.A and Australia as well as selected countries in the rest of the world.

How to perform the HBA™ assay

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Perform the assay at 20-30°C. Immediately before use, mix the semen and pipette a 7-10 µL drop onto the assay chamber.

Hba Assay


Immediately install the cover slip and avoid entrapping air bubbles. Incubate the chamber for at least 10 and not more than 20 minutes.

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In 10 minutes, all the mature spermatozoa with hyaluronan receptors contact and bind to the immobilized hyaluronan layer. Bound motile spermatozoa cease progressive movement but retain active tail beating.

Immature spermatozoa, lacking hyaluronan receptors, swim about freely.

Dead and non-motile spermatozoa show no tail movement.

HBA Assay


Calculate the percentage of hyaluronan-binding spermatozoa. The percentage of sperm binding to the hyaluronan layer is calculated as follows:

% Bound = 100 X (Bound Motile Sperm) / (Bound Motile Sperm + Unbound Motile Sperm)

The sample concentration should permit at least 30 motile spermatozoa to be counted. Acceptable assay precision will be achieved when the sum of bound plus unbound motile spermatozoa is greater than 200.

HBA index and ICSI treatment outcomes

ICSI patients with a low HBA index can benefit from hyaluronan-based sperm selection. Selection of mature spermatozoa, capable to bind to hyaluronan, increases the chance of picking spermatozoa with better DNA quality and can help to decrease miscarriage rates,6,7 and improve treatment outcomes in advance age patients,8 patients with previous unsuccessful ICSI attempts9 and patients with a low HBA index.6

After evaluation of the HBA index, PICSI® dishes or SpermSlow™ media can be used for selection of mature spermatozoa during ICSI procedures.

Learn More about hyaluronan-based sperm selection clinical efficiency

It was shown in a multicenter randomized trial that in couples where ≤65% of spermatozoa bound to hyaluronan, the selection of hyaluronan-bound spermatozoa for ICSI led to a statistically significant reduction in pregnancy loss rate compared to conventional ICSI.6

Outcome of a randomized controlled study, carried out in 10 IV clinics in USA6


HBA score positively correlates with fertilization, pregnancy and cleavage rates¹


Retrospective trial of 192 female patients and their male partners who underwent ICSI at Singapore General Hospital. *p-value <0.05


ORIGIO® Gradients and ORIGIO® Sperm Wash

Advanced media with innovative formulation for sperm preparation. Support sperm motility and function and protect spermatozoa from oxidative and osmotic stress during preparation.

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For mature sperm selection for ICSI. Recently published clinical data confirm sperm selection with PICSI dish can help to mitigate the negative effect of advanced maternal age

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For one step immobilization and selection of mature spermatozoa for ICSI without the use of PVP.

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Product codes

ProductOrder Code
HBA® slide: Package of 10 assaysBCT-HBA-10

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  1. Huszar G, Jakab A, Sakkas D, Ozenci CC, Cayli S, Delpiano E, Ozkavukcu S. Fertility testing and ICSI sperm selection by hyaluronic acid binding: clinical and genetic aspects. Reprod Biomed Online. 2007 May;14(5):650-63.
  2. Jakab A, Sakkas D, Delpiano E, Cayli S, Kovanci E, Ward D, Revelli A, Huszar G. Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies. Fertil Steril. 2005 Dec;84(6):1665-73.
  3. Parmegiani L, Cognigni GE, Bernardi S, Troilo E, Ciampaglia W, Filicori M. “Physiologic ICSI”: hyaluronic acid (HA) favors selection of spermatozoa without DNA fragmentation and with normal nucleus, resulting in improvement of embryo quality. Fertil Steril. 2010 Feb;93(2):598-604.
  4. Fen C.T.C., Lee S. N, Lim M. N, Yu S. L. Relationship between Sperm Hyaluronan-Binding Assay (HBA) Scores on Embryo Development, Fertilisation, and Pregnancy Rate in Patients Undergoing Intra-Cytoplasmic Sperm Injection (ICSI). Proceedings of Singapore Healthcare.2013.22.N2: 120-124.
  5. Nijs M, Creemers E, Cox A, Janssen M, Vanheusden E, Van der Elst J, Ombelet W. Relationship between hyaluronic acid binding assay and outcome in ART: a pilot study. Andrologia. 2010 Oct;42(5):291-6
  6. Worrilow KC, Eid S, Woodhouse D, Perloe M, Smith S, Witmyer J, Ivani K, Khoury C, Ball GD, Elliot T, Lieberman J. Use of hyaluronan in the selection of sperm for intracytoplasmic sperm injection (ICSI): significant improvement in clinical outcomes–multicenter, double-blinded and randomized controlled trial. Hum Reprod. 2013 Feb;28(2):306-14.
  7. Miller D, Pavitt S, Sharma V, Forbes G, Hooper R, Bhattacharya S, Kirkman-Brown J, Coomarasamy A, Lewis S, Cutting R, Brison D, Pacey A, West R, Brian K, Griffin D, Khalaf Y. Physiological, hyaluronan-selected intracytoplasmic sperm injection for infertility treatment (HABSelect): a parallel, two-group, randomised trial. Lancet. 2019 Feb 2;393(10170):416-422.
  8. West R, Coomarasamy A, Frew L, Hutton R, Kirkman-Brown J, Lawlor M, Lewis S, Partanen R, Payne-Dwyer A, Román-Montañana C, Torabi F, Tsagdi S, Miller D. Sperm selection with hyaluronic acid improved live birth outcomes among older couples and was connected to sperm DNA quality, potentially affecting all treatment outcomes. Hum Reprod. 2022 May 30;37(6):1106-1125.
  9. Scaruffi P, Bovis F, Casciano I, Maccarini E, De Leo C, Gazzo I, Massarotti C, Sozzi F, Stigliani S, Anserini P. Hyaluronic acid-sperm selection significantly improves the clinical outcome of couples with previous ICSI cycles failure. Andrology. 2022 May;10(4):677-685.



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