Microplate Reader versus Manual Water-Bath (37°C) Methods for Clot-Based Coagulation Testing: A Comparative Review

Dorathy Okpokam and Daniel Muoka

Background and Objective: Clot-based testing is a series of assays carried out to ascertain the functionality of the clotting proteins or enzymes. Over the years, the tilt tube method (grounded technique) using a water bath has been available in our local setting and in Nigeria as a whole. This review aimed to provide information on the usefulness of the microplate reader compared to the manual method (use of a water bath) in the clot-based testing method. Materials and Methods: This review adopted a narrative review approach to provide a comprehensive overview of the use of microplate readers and manual water bath (37°C) methods in clot-based coagulation testing. Relevant literature was systematically searched from major electronic databases, including PubMed, Scopus, Web of Science and Google Scholar. The search covered publications from January 2000 to December 2025, with emphasis on studies evaluating coagulation testing methodologies, laboratory automation and analytical performance of clot-based assays. Results: The reviewed literature indicates that manual water-bath methods are prone to variability arising from subjective clot endpoint determination, timing inconsistencies and potential temperature fluctuations that may influence coagulation enzyme activity. In contrast, microplate reader-based assays provide automated optical detection through absorbance, fluorescence, or luminescence measurements, thereby reducing observer bias and improving standardisation. Studies reviewed also demonstrated that microplate readers facilitate simultaneous analysis of multiple samples, improve workflow efficiency, reduce reagent consumption and enhance result reproducibility. These advantages make microplate reader technology a promising alternative to conventional manual methods for clot-based coagulation testing. Conclusion: From the review so far, the findings have shown that microplate reader-based coagulation testing offers significant advantages over manual water-bath methods in terms of automation, analytical consistency, sample throughput and reproducibility.

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