Development of the Dip Quick Wright-Giemsa Staining Methods for Improving Simplicity and Reducing Cost in Malaria Diagnosis

Wimol  Pararach; Audomsak  Churat; Rachadaporn  Benchawattananon

Authors

Wimol Pararach Student, Doctor of Philosophy in Biological Science, Department of Integrated Science, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand
Audomsak Churat Research assistant, Department of Integrated Science, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand
Rachadaporn Benchawattananon Associate Professor, Department of Integrated Science, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand

Keywords:

Wright-Giemsa staining, Malaria diagnosis

Abstract

Microscopy continues to be the benchmark for malaria diagnosis, however, the method seems not only complicate, but also time- and cost-consuming. This study presents the modified Wright-Giemsa staining technique by employing hospital tap water instead a commercial buffer, together with dip quick procedure. Comparative evaluation by 28 medical technicians of Loei Hospital, Thailand, for examining of 48,885 cases indicated more potential than the normally used method in malaria diagnosis and also other hematological identifications of blood cell types, such as red blood cells, leukocytes, and platelets. The developing dip quick method preserved time saving for 5 minutes per sample and 50% decrease of expenses, which were significant for applying as potentially alternative choice in urgent diagnostic situations wilderness areas.

References

Kolawole EO, Ayeni ET, Abolade SA, Ugwu SE, Awoyinka TB, Ofeh AS, et al. Malaria endemicity in Sub-Saharan Africa: Past and present issues in public health. Microbes Infect Dis. 2023;4(1):242-51.

Kogan F. Remote sensing for malaria: Monitoring and predicting malaria from operational satellites. Cham: Springer Nature; 2020.

Zheng J, Zhang D, Zhang S, Chen M, Guo Z, Guan S, et al. Global burden of malaria and neglected tropical diseases in children and adolescents, 1990-2019: A population-based cross-sectional study. J R Soc Med. 2025;118(3):82-96.

Moody A. Rapid diagnostic tests for malaria parasites. Clin Microbiol Rev. 2002;15(1):66-78.

Handayani L, Fitriani I. Review of Plasmodium vivax malaria in Indonesia and Malaysia: Epidemiology of the infection and the presence of the vector across time. Trop J Health Sci. 2025;32(2):47-56.

Akar SE, Dairo DM, Kehinde R, Erhabor T, Akar MO, Ntadom G, et al. A multitude of dyes but unsatisfactory staining: Physico-chemical profile and factors associated with staining quality of Romanowsky-type stains used for malaria microscopy in Plateau State, Nigeria. medRxiv. Forthcoming 2025.

Burton AG. Clinical atlas of small animal cytology and hematology. 2nd ed. New Jersey: John Wiley & Sons; 2024.

Gunawardena D, Priyankara I, Jayamanne H, Suresh S. Modified Giemsa stain: A solution to improve the quality of hypercellular bone marrow smears. Int J Lab Hematol. 2022;44(3):504-9.

Ani NV, Ogundunmade TP, Daniel D, Raheem KA, Odirichukwu EO, Osuagwuh UI. Comparative durability of common stains used for exfoliative vaginal cytology. Sahel J Vet Sci. 2021;18(2):9-16.

Oktiyani N, Muhlisin A, Roebiakto E, Norsiah W, Mahpolah M. Utilization of alternative buffer solutions for staining thin blood smears by the Giemsa, Wright stain and Romanowsky method. Trop Health Med Res. 2023;5(1):34-5.

Yue QF, Xiong B, Chen WX, Liu XY. Comparative study of the efficacy of Wright-Giemsa stain and Liu's stain in the detection of Auer rods in acute promyelocytic leukemia. Acta Histochem. 2014;116(6):1113-6.

Deepthi B, Prayaga AK, Rukmangadha N. Comparison of modified ultrafast Giemsa stain with the standard May-Grünwald Giemsa stain in FNAC of various organs. J Cytol. 2022;39(4):174-9.

Prole DL, Taylor CW. Identification of intracellular and plasma membrane calcium channel homologues in pathogenic parasites. PLoS One. 2011;6(10):e26218-33.

Piña R, Santos-Díaz AI, Orta-Salazar E, Aguilar-Vazquez AR, Mantellero CA, Acosta-Galeana I, et al. Ten approaches that improve immunostaining: A review of the latest advances for the optimization of immunofluorescence. Int J Mol Sci. 2022;23(3):1426.

Mebratie DY, Dagnaw GG. Review of immunohistochemistry techniques: Applications, current status, and future perspectives. Semin Diagn Pathol. 2024;41(3):154-60.

Tsutsumi Y. Pitfalls and caveats in applying chromogenic immunostaining to histopathological diagnosis. Cells. 2021;10(6):1501.

Sule R, Rivera G, Gomes AV. Western blotting (immunoblotting): History, theory, uses, protocol and problems. Biotechniques. 2023;75(3):99-114.

DuBois D, Binder L, Nelson B. Usefulness of the stool Wright's stain in the emergency department. J Emerg Med. 1988;6(6):483-6.

Shi Q, Wang C, Yang W, Ma X, Tang J, Zhang J, et al. Plasmodium falciparum transcription factor AP2-06B is mutated at high frequency in Southeast Asia but does not associate with drug resistance. Front Cell Infect Microbiol. 2025;14:1521152-64.

Singh K, Paul R, Chanu MD, Jayant S, Singh P, Naithani N, et al. Visual guide to clinical parasitology: A color atlas. Madhya Pradesh: Professional Publication Services; 2024.

Hazelton PR, Gelderblom HR. Electron microscopy for rapid diagnosis of emerging infectious agents. Emerg Infect Dis. 2003;9(3):294.

Onosakponome EO, Amadi CF, Bademosi A, Danagogo LS, Ikpeama RA. Comprehensive techniques in laboratory medicine: An in-depth guide. In: Sreenivasan A, Deepa PD, Chitambare K, Khan I, editors. Issues on health science. India: ZenToks Publication; 2024. p.21-65.

Chakraborty S. Democratizing nucleic acid-based molecular diagnostic tests for infectious diseases at resource-limited settings—from point of care to extreme point of care. Sens Diagn. 2024;3(4):536-61.

MacAulay S, Ellison AR, Kille P, Cable J. Moving towards improved surveillance and earlier diagnosis of aquatic pathogens: From traditional methods to emerging technologies. Rev Aquac. 2022;14(4):1813-29.

Fang W, Wu J, Cheng M, Zhu X, Du M, Chen C, et al. Diagnosis of invasive fungal infections: Challenges and recent developments. J Biomed Sci. 2023;30(1):42.

Locke D, Hoyt CC. Companion diagnostic requirements for spatial biology using multiplex immunofluorescence and multispectral imaging. Front Mol Biosci. 2023;10:1051491-517.

Dabbagh Moghaddam F, Rafiee M, Setayeshfar A, Moradi A, Esmaeili Y, Romana Bertani F, et al. Emerging trends and future prospects in microfluidic systems for prevention and diagnosis of infection pathogens. Adv Mater Technol. 2024;9(17):2400134.

Racey J. Development of novel turbidity calibration standards and methodologies using appropriate technology [MSc thesis]. Indiana: Purdue University; 2019.

Nielsen SS. Water hardness testing by complexometric determination of calcium. In: Food analysis laboratory manual. 3rd ed. Cham: Springer International Publishing; 2017. p.147-52.

Pagel JE, Qureshi AA, Young DM, Vlassoff LT. Comparison of four membrane filter methods for fecal coliform enumeration. Appl Environ Microbiol. 1982;43(4):787-93.

IBM Corporation. IBM SPSS statistics for Windows, version 29.0. New York: IBM Corporation; 2022.

Makler MT, Palmer CJ, Ager AL. A review of practical techniques for the diagnosis of malaria. Ann Trop Med Parasitol. 1998;92(4):419-34.

Milne LM, Kyi MS, Chiodini PL, Warhurst DC. Accuracy of routine laboratory diagnosis of malaria in the United Kingdom. J Clin Pathol. 1994;47(8):740-2.

Wongsrichanalai C, Barcus MJ, Muth S, Sutamihardja A, Wernsdorfer WH. A review of malaria diagnostic tools: Microscopy and rapid diagnostic test (RDT). Am J Trop Med Hyg. 2007;77(6 Suppl):119-27.

Funder J, Wieth JO. Chloride and hydrogen ion distribution between human red cells and plasma. Acta Physiol Scand. 1966;68(2):234-45.