Application of 18S rRNA Gene-Based and β-Giardin Molecular Markers for Early Detection of Giardia duodenalis Infection

Vol 9 No 1 (2026): Volume 09 Issue 01 January 2026

Article Date Published : 20 January 2026 | Page No.: 260-268 | Google Scholar | Crossref doi for this article

Abstract :

Giardiasis and other intestinal parasite infections are still prevalent and a public health concern, especially in areas with inadequate sanitation and hygiene. Giardia duodenalis infections can cause mild to severe symptoms, such as chronic diarrhea, malabsorption, and growth impairment in children, so early detection is essential. The commonly used fecal microscopy test has a low sensitivity, particularly for infections with low parasite loads or in the absence of symptoms. The development of molecular diagnostic methods based on the polymerase chain reaction (PCR) provides a more accurate and sensitive alternative by using specific genetic markers. This review of the literature looks at the roles of the 18S rRNA and β-giardin genes as molecular markers for the early detection of Giardia duodenalis infection. The review’s conclusions indicate that the 18S rRNA gene’s high sensitivity and robust sequence stability make it suitable for initial screening and epidemiological surveillance. In contrast, the β-giardin gene is useful for genotype analysis and diagnostic confirmation because of its higher specificity. The combination of these two genes may be the most effective diagnostic strategy to improve the accuracy of early Giardiasis detection in both clinical and epidemiological settings.

Keywords :

18S rRNA, Early Detection, Giardia duodenalis, molecular marker., PCR, β-giardin

References :

  1. Adam, R. (2021). Giardia duodenalis: Biology and Pathogenesis. Clinical Microbiology Reviews, 34. https://doi.org/10.1128/cmr.00024-19
  2. Ahmad, A., El-Kady, A., & Hassan, T. (2020). Genotyping of Giardia duodenalis in children in upper Egypt using assemblage- specific PCR technique. PLoS ONE, 15. https://doi.org/10.1371/journal.pone.0240119
  3. Cacciò, S., De Giacomo, M., & Pozio, E. (2002). Sequence analysis of the beta-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples. International Journal for Parasitology, 32 8, 1023–1030. https://doi.org/10.1016/s0020-7519(02)00068-1
  4. Cacciò, S. M., & Ryan, U. (2008a). Molecular epidemiology of giardiasis. Molecular and Biochemical Parasitology, 160(2), 75–80. https://doi.org/10.1016/j.molbiopara.2008.04.006
  5. Cacciò, S. M., & Ryan, U. (2008b). Molecular epidemiology of giardiasis. Molecular and Biochemical Parasitology, 160(2), 75–80. https://doi.org/10.1016/j.molbiopara.2008.04.006
  6. (2024). Giardiasis [Giardia duodenalis (syn. G. lamblia, G. intestinalis)]. https://www.cdc.gov/dpdx/giardiasis/index.html
  7. Elmahallawy, E. K., Gareh, A., Ghallab, M. M. I., Köster, P. C., Dashti, A., Aboelsoued, D., Toaleb, N. I., Alzaylaee, H., Gonzálvez, M., Saleh, A. A., Alhegaili, A. S., Eldehn, A. F., Hernández-Castro, C., Bailo, B., González-Barrio, D., & Carmena, D. (2024). Microscopy detection and molecular characterisation of Giardia duodenalis infection in outpatients seeking medical care in Egypt. Frontiers in Public Health, 12, 1377123. https://doi.org/10.3389/fpubh.2024.1377123
  8. Esmailikia, L., Ebrahimzade, E., Shayan, P., & Amininia, N. (2017). Detection of small number of Giardia in biological materials prepared from stray dogs. Acta Parasitologica, 62, 733–738. https://doi.org/10.1515/ap-2017-0088
  9. Faghiri, Z., & Widmer, G. (2011). A comparison of the Giardia lamblia trophozoite and cyst transcriptome using microarrays. BMC Microbiology, 11, 91. https://doi.org/10.1186/1471-2180-11-91
  10. Fu, Y., Dong, H., Bian, X., Qin, Z., Han, H., Lang, J., Zhang, J.-C., Zhao, G., Li, J., & Zhang, L. (2022). Molecular characterizations of Giardia duodenalis based on multilocus genotyping in sheep, goats, and beef cattle in Southwest Inner Mongolia, China. Parasite, 29. https://doi.org/10.1051/parasite/2022036
  11. Gatei, W., Greensill, J., Ashford, R., Cuevas, L., Parry, C., Cunliffe, N., Beeching, N., & Hart, C. (2003). Molecular Analysis of the 18S rRNA Gene of Cryptosporidium Parasites from Patients with or without Human Immunodeficiency Virus Infections Living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. Journal of Clinical Microbiology, 41, 1458–1462. https://doi.org/10.1128/jcm.41.4.1458-1462.2003
  12. Gutiérrez, L., & Bartelt, L. (2024). Current Understanding of Giardia lamblia and Pathogenesis of Stunting and Cognitive Deficits in Children from Low- and Middle-Income Countries. Current Tropical Medicine Reports, 11(1), 28–39. https://doi.org/10.1007/s40475-024-00314-2
  13. Guy, R. A., Xiao, C., & Horgen, P. A. (2004). Real-time PCR assay for detection and genotype differentiation of Giardia lamblia in stool specimens. Journal of Clinical Microbiology, 42(7), 3317–3320. https://doi.org/10.1128/JCM.42.7.3317-3320.2004
  14. Hajare, S. T., Chekol, Y., & Chauhan, N. M. (2022). Assessment of prevalence of Giardia lamblia infection and its associated factors among government elementary school children from Sidama zone, SNNPR, Ethiopia. PloS One, 17(3), e0264812. https://doi.org/10.1371/journal.pone.0264812
  15. Hijjawi, N., Yang, R., Hatmal, M., Yassin, Y., Mharib, T., Mukbel, R., Mahmoud, S. A., Al-Shudifat, A.-E., & Ryan, U. (2018). Comparison of ELISA, nested PCR and sequencing and a novel qPCR for detection of Giardia isolates from Jordan. Experimental Parasitology, 185, 23–28. https://doi.org/10.1016/j.exppara.2018.01.011
  16. Hooshyar, H., Rostamkhani, P., Arbabi, M., & Delavari, M. (2019). Giardia lamblia infection: review of current diagnostic strategies. Gastroenterology and Hepatology from Bed to Bench, 12(1), 3–12.
  17. Hussain, S., Sharif, A., Bashir, F., Ali, S., Javid, A., Hussain, A. I., Ghafoor, A., Alshehri, M. A., Naeem, A., Naeem, E., & Amjad, M. (2025). Polymerase Chain Reaction: A Toolbox for Molecular Discovery. Molecular Biotechnology. https://doi.org/10.1007/s12033-025-01390-z
  18. Jenkins, M., O’brien, C., Murphy, C., Schwarz, R., Miska, K., Rosenthal, B., & Trout, J. (2009). Antibodies to the Ventral Disc Protein δ-giardin Prevent in Vitro Binding of Giardia lamblia Trophozoites. 95, 895–899. https://doi.org/10.1645/ge-1851r.1
  19. Jothikumar, N., Murphy, J. L., & Hill, V. R. (2021). Detection and identification of Giardia species using real-time PCR and Journal of Microbiological Methods, 189, 106279. https://doi.org/10.1016/j.mimet.2021.106279
  20. Kounosu, A., Murase, K., Yoshida, A., Maruyama, H., & Kikuchi, T. (2019). Improved 18S and 28S rDNA primer sets for NGS-based parasite detection. Scientific Reports, 9. https://doi.org/10.1038/s41598-019-52422-z
  21. Kralik, P., & Ricchi, M. (2017). A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything. Frontiers in Microbiology, 8. https://doi.org/10.3389/fmicb.2017.00108
  22. Kumagai, H., & Furusawa, H. (2024). Real-Time Monitoring of a Nucleic Acid Amplification Reaction Using a Mass Sensor Based on a Quartz-Crystal Microbalance. Biosensors, 14. https://doi.org/10.3390/bios14040155
  23. Kuthyar, S., Kowalewski, M., Seabolt, M., Roellig, D., & Gillespie, T. (2021). Molecular characterization of Giardia duodenalis and evidence for cross-species transmission in Northern Argentina. Transboundary and Emerging Diseases. https://doi.org/10.1111/tbed.14220
  24. Lee, H., & Kwak, D. (2023). Molecular detection and assemblage analysis of the intestinal protozoan Giardia duodenalis in wild boars in Korea. Frontiers in Veterinary Science, 10, 1139060. https://doi.org/10.3389/fvets.2023.1139060
  25. Lehto, K.-M., Fan, Y.-M., Oikarinen, S., Nurminen, N., Hallamaa, L., Juuti, R., Mangani, C., Maleta, K., Hyöty, H., & Ashorn, P. (2019). Presence of Giardia lamblia in stools of six- to 18-month old asymptomatic Malawians is associated with children’s growth failure. Acta Paediatrica (Oslo, Norway : 1992), 108(10), 1833–1840. https://doi.org/10.1111/apa.14832
  26. Leung, A., Leung, A., Wong, A., Sergi, C., & Kamsites, J. (2019). Giardiasis : An overview. Recent Patents on Inflammation & Allergy Drug Discovery. https://doi.org/10.2174/1872213×13666190618124901
  27. Lourenço, D., Andrade, I. da S., Terra, L. L., Guimarães, P. R., Zingali, R. B., & de Souza, W. (2012). Proteomic analysis of the ventral disc of Giardia lamblia. BMC Research Notes, 5, 41. https://doi.org/10.1186/1756-0500-5-41
  28. Mala, S., & Hamza, D. (2016). Real-time PCR Assay for Diagnosis of Eukaryotic Peptide Chain Release Factor Subunit 1 Gene in the Stages of Giardia duodenalis. Journal of Babylon University, 24, 1234–1245. https://consensus.app/papers/realtime-pcr-assay-for-diagnosis-of-eukaryotic-peptide-mala-hamza/7de7f0969b7f5a8c967aff6b89ae0b8c/
  29. Messa, A., Köster, P., Garrine, M., Gilchrist, C., Bartelt, L., Nhampossa, T., Massora, S., Kotloff, K., Levine, M., Alonso, P., Carmena, D., & Mandomando, I. (2021). Molecular diversity of Giardia duodenalis in children under 5 years from the Manhiça district, Southern Mozambique enrolled in a matched case-control study on the aetiology of diarrhoea. PLoS Neglected Tropical Diseases, 15. https://doi.org/10.1371/journal.pntd.0008987
  30. Molina-Gonzalez, S., Bhattacharyya, T., AlShehri, H., Poulton, K., Allen, S., Miles, M., Arianitwe, M., Tukahebwa, E., Webster, B., Stothard, R., & Bustinduy, A. (2020). Application of a recombinase polymerase amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda. Parasites & Vectors, 13. https://doi.org/10.1186/s13071-020-04168-1
  31. Monteiro, L., Bonnemaison, D., Vekris, A., Petry, K. G., Bonnet, J., Vidal, R., Cabrita, J., & Mégraud, F. (1997). Complex polysaccharides as PCR inhibitors in feces: Helicobacter pylori model. Journal of Clinical Microbiology, 35(4), 995–998. https://doi.org/10.1128/jcm.35.4.995-998.1997
  32. Moreno, Y., Moreno-Mesonero, L., Amorós, I., Pérez, R., Morillo, J., & Alonso, J. (2018). Multiple identification of most important waterborne protozoa in surface water used for irrigation purposes by 18S rRNA amplicon-based metagenomics. International Journal of Hygiene and Environmental Health, 221 1, 102–111. https://doi.org/10.1016/j.ijheh.2017.10.008
  33. Morris, A., Robinson, G., Chalmers, R., Caccio, S., & Connor, T. (2025). Parapipe: a Pipeline for Handling Parasite NGS Datasets and its Application to Cryptosporidium. BioRxiv. https://doi.org/10.1101/2025.01.16.633364
  34. Nantavisai, K., Mungthin, M., Tan-ariya, P., Rangsin, R., Naaglor, T., & Leelayoova, S. (2007). Evaluation of the sensitivities of DNA extraction and PCR methods for detection of Giardia duodenalis in stool specimens. Journal of Clinical Microbiology, 45(2), 581–583. https://doi.org/10.1128/JCM.01823-06
  35. Narayanan, J., Murphy, J., & Hill, V. (2021). Detection and identification of Giardia species using real-time PCR and sequencing. Journal of Microbiological Methods, 106279. https://doi.org/10.1016/j.mimet.2021.106279
  36. Ng, C., Gilchrist, C., Lane, A., Roy, S., Haque, R., & Houpt, E. (2005). Multiplex Real-Time PCR Assay Using Scorpion Probes and DNA Capture for Genotype-Specific Detection of Giardia lamblia on Fecal Samples. Journal of Clinical Microbiology, 43, 1256–1260. https://doi.org/10.1128/jcm.43.3.1256-1260.2005
  37. Pallant, L., Barutzki, D., Schaper, R., & Thompson, R. (2015). The epidemiology of infections with Giardia species and genotypes in well cared for dogs and cats in Germany. Parasites & Vectors, 8. https://doi.org/10.1186/s13071-014-0615-2
  38. Pisarev, A., Kolupaeva, V., Yusupov, M., Hellen, C., & Pestova, T. (2008). Ribosomal position and contacts of mRNA in eukaryotic translation initiation complexes. The EMBO Journal, 27. https://doi.org/10.1038/emboj.2008.90
  39. Pouryousef, A., Fararouei, M., & Sarkari, B. (2023). Antigen-Based Diagnosis of Human Giardiasis: A Systematic Review and Meta-Analysis. Iranian Journal of Parasitology, 18(2), 140–154. https://doi.org/10.18502/ijpa.v18i2.13180
  40. Prasertbun, R., Sukthana, Y., & Popruk, S. (2012). Real-time PCR: Benefits for Detection of Mild and Asymptomatic Giardia Tropical Medicine and Health, 40(2), 31–35. https://doi.org/10.2149/tmh.2012-08
  41. Quan, P., Sauzade, M., & Brouzes, E. (2018). dPCR: A Technology Review. Sensors (Basel, Switzerland), 18. https://doi.org/10.3390/s18041271
  42. Rafiei, A., Baghlaninezhad, R., Köster, P., Bailo, B., De Mingo, M. H., Carmena, D., Panabad, E., & Beiromvand, M. (2020). Multilocus genotyping of Giardia duodenalis in Southwestern Iran. A community survey. PLoS ONE, 15. https://doi.org/10.1371/journal.pone.0228317
  43. Ryan, U., & Cacciò, S. M. (2013). Zoonotic potential of Giardia. International Journal for Parasitology, 43(12–13), 943–956. https://doi.org/10.1016/j.ijpara.2013.06.001
  44. Savioli, L., Smith, H., & Thompson, A. (2006). Giardia and Cryptosporidium join the “Neglected Diseases Initiative”. Trends in Parasitology, 22(5), 203–208. https://doi.org/10.1016/j.pt.2006.02.015
  45. Shamsi, L., Shadfar, F., Bahramdoost, Z., Pouryousef, A., Asghari, A., & Maleki, F. (2025). Diagnostic Accuracy of Various Genetic Markers (tpi, gdh, and bg) for Prevalence and Genotyping of Giardia duodenalis in Human Samples: A Comparative Global Systematic Review and Meta-Analysis. Acta Parasitologica, 70. https://doi.org/10.1007/s11686-025-01101-3
  46. Shumanskaya, S., Zhyzneuskaya, A., Grigorieva, E., Dronina, A., & Fomina, E. (2025). Pathogenetic features of Giardia lamblia, achievements and prospects for genotyping the protozoan. Health and Ecology Issues. https://doi.org/10.51523/2708-6011.2024-21-4-01
  47. Simsek, Z., Zeyrek, F. Y., & Kurcer, M. A. (2004). Effect of Giardia infection on growth and psychomotor development of children aged 0-5 years. Journal of Tropical Pediatrics, 50(2), 90–93. https://doi.org/10.1093/tropej/50.2.90
  48. Stephen, T., Tobin, E. H., & Juergens, A. L. (2025). Giardiasis. StatPearls Publishing. https://www.ncbi.nlm.nih.gov/sites/books/NBK513239/
  49. Tejan, N., Siddiqui, T., Singh, P., Ghoshal, U. C., & Ghoshal, U. (2023). Trends and clinico-epidemiological profile of patients with Giardiasis: Five-year experience from a university hospital of northern India. Journal of Family Medicine and Primary Care, 12(10), 2451–2455. https://doi.org/10.4103/jfmpc.jfmpc_2337_22
  50. Trevethan, R. (2017). Sensitivity, Specificity, and Predictive Values: Foundations, Pliabilities, and Pitfalls in Research and Practice. Frontiers in Public Health, 5, 307. https://doi.org/10.3389/fpubh.2017.00307
  51. Uchôa, F. F. de M., Sudré, A. P., Macieira, D. de B., & Almosny, N. R. P. (2017). The influence of serial fecal sampling on the diagnosis of giardiasis in humans, dogs, and cats. Revista Do Instituto de Medicina Tropical de Sao Paulo, 59, e61. https://doi.org/10.1590/S1678-9946201759061
  52. Veeken, L., Schwalb, A., Horton, K., Koesoemadinata, R., Alisjahbana, B., Van Crevel, R., & Houben, R. (2025). Mind the Clinic-Community Gap: Re-evaluation of Test Performance and False Positive Results in Community-Wide Tuberculosis Screening. The Journal of Infectious Diseases, 232. https://doi.org/10.1093/infdis/jiaf268
  53. Verweij, J. J., Schinkel, J., Laeijendecker, D., van Rooyen, M. A. A., van Lieshout, L., & Polderman, A. M. (2003). Real-time PCR for the detection of Giardia lamblia. Molecular and Cellular Probes, 17(5), 223–225. https://doi.org/10.1016/s0890-8508(03)00057-4
  54. Vicente, B., Freitas, A. De, Freitas, M., & Midlej, V. (2024). Systematic Review of Diagnostic Approaches for Human Giardiasis: Unveiling Optimal Strategies. Diagnostics (Basel, Switzerland), 14(4). https://doi.org/10.3390/diagnostics14040364
  1. Volotão, A. C., Costa-Macedo, L. M., Haddad, F. S. M., Brandão, A., Peralta, J. M., & Fernandes, O. (2007). Genotyping of Giardia duodenalis from human and animal samples from Brazil using beta-giardin gene: a phylogenetic analysis. Acta Tropica, 102(1), 10–19. https://doi.org/10.1016/j.actatropica.2007.02.010
  2. Wang, H., Yang, H., Li, C., Chen, M., Wang, X., Zhang, X., Gong, P., Zhang, N., Zhang, X., Li, J., & Li, X. (2025). β-Giardin as an Immunomagnetic Enrichment Target for Multi-Host Detection of Giardia duodenalis Cysts. Pathogens (Basel, Switzerland), 14(9). https://doi.org/10.3390/pathogens14090918
  3. Weinreich, F., Hahn, A., Eberhardt, K. A., Kann, S., Feldt, T., Sarfo, F. S., Di Cristanziano, V., Frickmann, H., & Loderstädt, U. (2022). Comparative Evaluation of Real-Time Screening PCR Assays for Giardia duodenalis and of Assays Discriminating the Assemblages A and B. Microorganisms, 10(7). https://doi.org/10.3390/microorganisms10071310
  4. Wu, Y., Yao, L., Chen, H., Zhang, W., Jiang, Y.-Y., Yang, F., Liu, A., & Shen, Y. (2022). Giardia duodenalis in patients with diarrhea and various animals in northeastern China: prevalence and multilocus genetic characterization. Parasites & Vectors, 15. https://doi.org/10.1186/s13071-022-05269-9
  5. Xiao, L., & Feng, Y. (2017). Molecular epidemiologic tools for waterborne pathogens Cryptosporidium spp. and Giardia duodenalis. Food and Waterborne Parasitology, 8–9, 14–32. https://doi.org/10.1016/j.fawpar.2017.09.002

How to Cite :

Application of 18S rRNA Gene-Based and β-Giardin Molecular Markers for Early Detection of Giardia duodenalis Infection. Nabila Aulia Safitri, Betta Kurniawan, Wiwi Febriani, Jhons Fatriyadi Suwandi, 9(1), 260-268. Retrieved from https://ijcsrr.org/single-view/?id=25184&pid=24992

Most read articles by the same author(s)

Nabila Aulia Safitri, Betta Kurniawan, Wiwi Febriani, Jhons Fatriyadi Suwandi,

View

147

Citation

Downloads

Article Details