Comparison of microscopy, ELISA, and real-time PCR for detection of Giardia intestinalis in human stool specimens

Comparison of microscopy, ELISA, and real-time PCR for detection of Giardia intestinalis in human stool specimens

Background/aim: This study included patients who had digestive system complaints between August 2015 and October 2015. The research was designed to compare conventional microscopy with an antigen detection ELISA kit and the TaqMan-based real-time PCR (RT-PCR) technique for detection of Giardia intestinalis in human stool specimens. Materials and methods: Samples were concentrated by formalin-ether sedimentation technique and microscopic examinations were carried out on wet mount slides. A commercially available ELISA kit (Giardia CELISA, Cellabs, Brookvale, Australia) was used for immunoassay. DNA was extracted from fecal samples of about 200 mg using the QIAamp Fast DNA Stool Mini Kit (QIAGEN, Hilden, Germany) and the LightCyclerNano system (Roche Diagnostics, Mannheim, Germany) was used for the TaqMan-based RT-PCR assay. Results: A total of 94 stool samples, 38 of them diagnosed positive (40.4%) and 56 of them diagnosed negative by microscopy, were selected for evaluation by antigen detection and molecular assays. The prevalence of G. intestinalis infection was found as 46.8% (n: 44) and 79.8% (n: 75) by ELISA and RT-PCR, respectively. RT-PCR revealed by far the highest positivity rate compared to the other two methods. The difference between these methods was found to be statistically significant (P < 0.05). In comparison to PCR, the sensitivity and specificity of microscopy and ELISA were 50.7% and 100% and 53.3% and 79%, respectively. Conclusion: RT-PCR seems to be much more sensitive and beneficial for rapid and accurate diagnosis of G. intestinalis in human stools

___

  • 1. Adam RDA. Biology of Giardia intestinalis. Clin Microbiol Rev 2001; 14: 447-475.
  • 2. WHO. The World Health Report. Fighting Disease, Fostering Development. Geneva, Switzerland: World Health Organization; 1996.
  • 3. Hill DR. Giardia intestinalis. In: Mandell GL, Bennett JE, Dolin R, editors. Mandell, Douglas, and Bennett’s Principles and Practice of Infectious Diseases. 8th ed. New York, NY, USA: Churchill Livingstone; 2010. pp. 3527-3534.
  • 4. Lane S, Lloyd D. Current trends in research into the waterborne parasite Giardia. Crit Rev Microbiol 2002; 28: 123-147.
  • 5. Ortega YR, Adam RD. Giardia: overview and update. Clin Infect Dis 1997; 25: 545-550.
  • 6. Garcia LS. Diagnostic Medical Parasitology. 5th ed. Washington, DC, USA: ASM Press; 2007.
  • 7. Johnston SP, Ballard MM, Beach MJ, Causer L, Wilkins PP. Evaluation of three commercial assays for the detection of Giardia and Cryptosporidium organisms in fecal specimens. J Clin Microbiol 2003; 41: 623-626.
  • 8. Babaei Z, Oormazdi H, Rezaie S, Rezaeian M, Razmjou E. Giardia intestinalis: DNA extraction approaches to improve PCR results. Exp Parasitol 2011; 128: 159-162.
  • 9. McGlade TR, Robertson ID, Elliot AD, Read C, Thompson RC. Gastrointestinal parasites of domestic cats in Perth, Western Australia. Vet Parasitol 2003; 117: 251-262.
  • 10. Klein D. Quantification using real–time PCR technology: applications and limitations. Trends Mol Med 2002; 8: 257-260.
  • 11. Caccio SM, Sprong H. Epidemiology of giardiasis in humans. In: Lujan HD, Svard S, editors. Giardia: A Model Organism. Vienna, Austria: Springer-Verlag; 2011. pp. 17-25.
  • 12. CDC. Division of Parasitic Diseases Program Review. National Center for Infectious Diseases. Atlanta, GA, USA: Centers for Disease Control; 1995.
  • 13. Marshall MM, Naumovitz D, Ortega Y, Sterling CR. Waterborne protozoan pathogens. Clin Microbiol Rev 1997; 10: 67-85.
  • 14. Balcioglu C, Limoncu E, Ertan P, Yereli K, Ozbilgin A, Onag A. Incidence of giardiasis among siblings in Turkey. Pediatr Int 2003; 45: 311-313.
  • 15. Dutt P, Mehta S, Vinayak VK. Enzyme linked immunosorbent assay for copro-diagnosis of giardiasis and characterization of a specific Giardia intestinalis antigen in stools. J Med Microbiol 1991; 34: 271-275.
  • 16. Clinical and Laboratory Standards Institute. Procedures for the Recovery and Identification of Parasites from the Intestinal Tract; Approved Guideline. CLSI Document M28-A2. 2nd ed. Wayne, PA, USA: Clinical Laboratory Standards Institute; 2005.
  • 17. Hanson KL, Cartwright CP. Use of an enzyme immunoassay does not eliminate the need to analyze multiple stool specimens for sensitive detection of Giardia intestinalis. J Clin Microbiol 2001; 39: 474-477.
  • 18. El-Nahas HA, Salem DA, El-Henawy AA, El-Nimr HI, AbdelGhaffar HA, El-Meadawy AM. Giardia diagnostic methods in human fecal samples: a comparative study. Cytometry B Clin Cytom 2013; 84: 44-49.
  • 19. Al-Saeed AT, Issa SH. Detection of Giardia intestinalis antigen in stool specimens using enzyme-linked immunosorbent assay. East Mediterr Health J 2010; 6: 362-364.
  • 20. Calderaro A, Gorrini C, Montecchini S, Peruzzi S, Piccolo G, Rossi S, Gargiulo F, Manca N, Dettori G, Chezzi C. Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis. Diagn Microbiol Infect Dis 2010; 66: 261-267.
  • 21. Addiss DG, Mathews HM, Stewart JM, Wahlquist SP, Williams RM, Finton RJ, Spencer HC, Juranek DD. Evaluation of a commercially available Enzyme-linked immunosorbent assay for Giardia intestinalis antigen in stool. J Clin Microbiol 1991; 29: 1137-1142.
  • 22. Jelinek T, Neifer S. Detection of Giardia intestinalis stool samples: a comparison of two enzyme-linked immunosorbent assays. F1000Research 2013; 2: 39-44.
  • 23. Aldeen WE, Carroll K, Robison A, Morrison M, Hale D. Comparison of nine commercially available enzyme-linked immunosorbent assays for detection of Giardia intestinalis in fecal specimens. J Clin Microbiol 1998; 36: 1338-1340.
  • 24. Schunk M, Jelinek T, Wetzel K, Nothdurft HD. Detection of Giardia intestinalis and Entamoeba histolytica in stool samples by two enzyme immunoassays. Eur J Clin Microbiol Infect Dis 2001; 20: 389-391.
  • 25. Maraha B, Buiting AGM. Evaluation of four enzyme immunoassays for the detection of Giardia intestinalis antigen in stool specimens. Eur J Clin Microbiol Infect Dis 2000; 19: 485-487.
  • 26. Stark D, Al-Qassab SE, Barratt JLN, Stanley K, Roberts T, Marriott D, Harkness J, Ellis JT. Evaluation of multiplex tandem real–time PCR for detection of Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis in clinical stool samples. J Clin Microbiol 2011; 49: 257-262.
  • 27. Verweij JJ, Schinkel J, Laeijendecker D, van Rooyen MA, van Lieshout L, Polderman AM. Real-time PCR for the detection of Giardia intestinalis. Mol Cell Probes 2003; 17: 223-225.
  • 28. Schuurman T, Lankamp P, van Belkum A, Kooistra-Smid M, van Zwet A. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia intestinalis in human stool specimens. Clin Microbiol Infect 2007; 13: 1186- 1191.
Turkish Journal of Medical Sciences-Cover
  • ISSN: 1300-0144
  • Yayın Aralığı: 6
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Mehmet CEYHAN, İnci YILDIRIM, Hasan TEZER, İlker DEVRİM, Emmanuel FEROLDI

The knowledge and high seroprevalence of hepatitis A in a high-risk group (agricultural reproductive-aged women) in the southeastern region of Turkey

Nebiye YENTÜR DONİ, Gülcan GÜRSES, Zeynep ŞİMŞEK, Fadile YILDIZ ZEYREK, Muhsin AKBABA

Oğuz Reşat SİPAHİ, SERHAT UYSAL, Sabire Şöhret AYDEMİR, Hüsnü PULLUKÇU, MELTEM TAŞBAKAN, ALPER TÜNGER, FATMA FERİHA ÇİLLİ, TANSU YAMAZHAN, BİLGİN ARDA, Hilal SİPAHİ, Sercan ULUSOY

Differences in leptin, ghrelin, and glucagon-like peptide-1 levels between religious fasting and normal fasting

Özge TELCİ ÇAKLILI, Aytekin OĞUZ, Banu MESÇİ, Banu İŞBİLEN, Gülcan YAVUZ, Selcan TÜLÜ

Decreased oxidative stress may contribute to the disease process in placenta accreta

Özcan EREL, Ali Turhan ÇAĞLAR, Aykan YÜCEL, Sibel ÖZLER, Merve ERGİN, Nuri DANIŞMAN, Dilek UYGUR, Efser ÖZTAŞ, Başak GÜMÜŞ GÜLER

Relationship of paratracheal lymph nodes with the progression of chronic autoimmune thyroiditis: 5-year follow-up results

Dilek BERKER, Gülhan AKBABA, Serhat IŞIK, Yasemin TÜTÜNCÜ, Ferit Kerim KÜÇÜKLER, Narin NASIROĞLU İMGA, Serdar GÜLER

Relationship between selected micronutrient deficiencies and oxidative stress biomarkers in diabetes mellitus patients with foot ulcers in Ibadan, Nigeria

Elizabeth Bosede BOLAJOKO, Olubayo Michael AKINOSUN, John ANETOR, Kensese Sontin MOSSANDA

Choel LEE, Jae-yoon CHUNG, Myeongjong LEE

A fully liquid DTaP-IPV-HB-PRP-T hexavalent vaccine for primary and booster vaccination of healthy Turkish infants and toddlers

İnci YILDIRIM, Mehmet CEYHAN, Hasan TEZER, İlker DEVRİM, Emmanuel FEROLDI

Güler Duru AŞİRET, Gülcan BAĞÇİVAN, Münevver ÖZCAN, Beste Başak ERÖKSÜZ, Nalan AKBAYRAK, Nuran AKDEMİR