Lab Med Online.  2020 Jul;10(3):221-226. 10.3343/lmo.2020.10.3.221.

Detection of Intestinal Protozoa in Korean Patients Using BD MAX Enteric Parasite Panel and Seegene Allplex Gastrointestinal Parasite Assay

• Kim B
• Kim HS
• Kim JJ
• Park YJ
• Kim D
• Yong D

• KMID: 2504132
• DOI: http://doi.org/10.3343/lmo.2020.10.3.221

Abstract

Background
Intestinal protozoan infection is one of the main causes of gastrointestinal diseases. Protozoa are usually detected by direct smear microscopy, concentration techniques, or special stains; however, these techniques are labor-intensive and require well-trained technicians. Therefore, molecular techniques involving polymerase chain reaction (PCR) have been developed to satisfy the need for unbiased and rapid analytical methods with high sensitivity and specificity. In this study, the BD MAX^TM Enteric Parasite Panel (EPP) (Becton, Dickinson and Company, USA), designed to detect Cryptosporidium parvum and/or hominis, Giardia lamblia, and Entamoeba histolytica, and the Allplex^TM Gastrointestinal Parasite Assays (AGPA) (Seegene Inc., Korea), designed to detect Cryptosporidium species, G. lamblia, E. histolytica, Blastocystis hominis, Dientamoeba fragilis, and Cyclospora cayetanensis were compared to determine whether any of these assays could become a useful tool for detecting intestinal protozoan infections in Korea.
Methods
We investigated 295 fecal samples using EPP and AGPA. Then we confirmed the positive results with the conventional and nested PCR. Consistent detection by conventional PCR, nested PCR, and one of the multiplex panels was considered “true positive.”
Results
Out of 295 samples, 17 were true positives for B. hominis and 2 were true positives for E. histolytica. EPP detected parasites in only two samples owing to its design; however, its true positive detection rate was 100% (2/2). AGPA detected parasites in 24 samples with 79.2% (19/24) true positives.
Conclusions
The incidence of protozoan, especially B. hominis, infection may be more prevalent than expected. AGPA could be an effective tool for screening protozoan infections.

Keyword

Parasite infection; Real-time PCR; Blastocystis hominis

Reference

1. Fletcher SM, Stark D, Harkness J, Ellis J. 2012; Enteric protozoa in the developed world: a public health perspective. Clin Microbiol Rev. 25:420–49. DOI: 10.1128/CMR.05038-11. PMID: 22763633. PMCID: PMC3416492.

2. Won EJ, Kim SH, Kee SJ, Shin JH, Suh SP, Chai JY, et al. 2016; Multiplex real-time PCR assay targeting eight parasites customized to the Korean population: Potential use for detection in diarrheal stool samples from gastroenteritis patients. PLoS One. 11:e0166957. DOI: 10.1371/journal.pone.0166957. PMID: 27861635. PMCID: PMC5115832.

3. Autier B, Belaz S, Razakandrainibe R, Gangneux JP, Robert-Gangneux F. 2018; Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa. Parasite. 25:48. DOI: 10.1051/parasite/2018049. PMID: 30230444. PMCID: PMC6144649.

4. Ocaña-Losada C, Cuenca-Gómez JA, Cabezas-Fernández MT, Vázquez-Villegas J, Soriano-Pérez MJ, Cabeza-Barrera I, et al. 2018; Clinical and epidemiological characteristics of intestinal parasite infection by Blastocystis hominis. Rev Clin Esp. 218:115–20. DOI: 10.1016/j.rce.2018.01.003. PMID: 29455921.

5. Mergani MH, Mohammed MA, Khan N, Bano M, Khan AH. 2014; Detection of intestinal protozoa by using different methods. Dent Med Res. 2:28–32. DOI: 10.4103/2348-1471.143326.

6. Ragavan AD, Govind SK. 2015; Modified fields' stain: ideal to differentiate Dientamoeba fragilis and Blastocystis sp. Parasitol Res. 114:1163–6. DOI: 10.1007/s00436-014-4296-8. PMID: 25614298.

7. McHardy IH, Wu M, Shimizu-Cohen R, Couturier MR, Humphries RM. 2014; Detection of intestinal protozoa in the clinical laboratory. J Clin Microbiol. 52:712–20. DOI: 10.1128/JCM.02877-13. PMID: 24197877. PMCID: PMC3957779.

8. Van den Bossche D, Cnops L, Verschueren J, Van Esbroeck M. 2015; Comparison of four rapid diagnostic tests, ELISA, microscopy and PCR for the detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in feces. J Microbiol Methods. 110:78–84. DOI: 10.1016/j.mimet.2015.01.016. PMID: 25615719.

9. Laude A, Valot S, Desoubeaux G, Argy N, Nourrisson C, Pomares C, et al. 2016; Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identi cation of Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review. Clin Microbiol Infect. 22:190. e1-8. DOI: 10.1016/j.cmi.2015.10.019. PMID: 26548509.

10. Madison-Antenucci S, Relich RF, Doyle L, Espina N, Fuller D, Karchmer T, et al. 2016; Multicenter evaluation of BD Max Enteric Parasite Real-Time PCR Assay for detection of Giardia duodenalis, Cryptosporidium hominis, Cryptosporidium parvum, and Entamoeba histolytica. J Clin Microbiol. 54:2681–8. DOI: 10.1128/JCM.00765-16. PMID: 27535690. PMCID: PMC5078543.

11. Verweij JJ. 2014; Application of PCR-based methods for diagnosis of intestinal parasitic infections in the clinical laboratory. Parasitology. 141:1863–72. DOI: 10.1017/S0031182014000419. PMID: 24780241.

12. Adeyemo FE, Singh G, Reddy P, Stenström TA. 2018; Methods for the detection of Cryptosporidium and Giardia: From microscopy to nucleic acid based tools in clinical and environmental regimes. Acta Trop. 184:15–28. DOI: 10.1016/j.actatropica.2018.01.011. PMID: 29395034.

13. Parčina M, Reiter-Owona I, Mockenhaupt FP, Vojvoda V, Gahutu JB, Hoerauf A, et al. 2018; Highly sensitive and specific detection of Giardia duodenalis, Entamoeba histolytica, and Cryptosporidium spp. in human stool samples by the BD MAX™ Enteric Parasite Panel. Parasitol Res. 117:447–51. DOI: 10.1007/s00436-017-5720-7. PMID: 29260297.

14. Perry MD, Corden SA, Lewis White P. 2017; Evaluation of the BD MAX Enteric Parasite Panel for the detection of Cryptosporidium parvum/hominis, Giardia duodenalis and Entamoeba histolytica. J Med Microbiol. 66:1118–23. DOI: 10.1099/jmm.0.000558. PMID: 28792380.

15. Yoo J, Park J, Lee HK, Yu JK, Lee GD, Park KG, et al. 2019; Comparative evaluation of Seegene Allplex Gastrointestinal, Luminex xTAG Gastrointestinal Pathogen Panel, and BD MAX Enteric Assays for detection of gastrointestinal pathogens in clinical stool specimens. Arch Pathol Lab Med. 143:999–1005. DOI: 10.5858/arpa.2018-0002-OA. PMID: 30763118.

16. Tan KS. 2008; New insights on classification, identification, and clinical relevance of Blastocystis spp. Clin Microbiol Rev. 21:639–65. DOI: 10.1128/CMR.00022-08. PMID: 18854485. PMCID: PMC2570156.

17. Roberts T, Stark D, Harkness J, Ellis J. 2013; Subtype distribution of Blastocystis isolates from a variety of animals from New South Wales, Australia. Vet Parasitol. 196:85–9. DOI: 10.1016/j.vetpar.2013.01.011. PMID: 23398989.

18. Clark CG. 2000; Cryptic genetic variation in parasitic protozoa. J Med Microbiol. 49:489–91. DOI: 10.1099/0022-1317-49-6-489. PMID: 10847200.

19. Stenzel DJ, Boreham PF. 1996; Blastocystis hominis revisited. Clin Microbiol Rev. 9:563–84. DOI: 10.1128/CMR.9.4.563. PMID: 8894352. PMCID: PMC172910.

20. Rajah Salim H, Suresh Kumar G, Vellayan S, Mak JW, Khairul Anuar A, Init I, et al. 1999; Blastocystis in animal handlers. Parasitol Res. 85:1032–3. DOI: 10.1007/s004360050677. PMID: 10599928.

21. Kurt Ö, Doğruman Al F, Tanyüksel M. 2016; Eradication of Blastocystis in humans: Really necessary for all? Parasitol Int. 65:797–801. DOI: 10.1016/j.parint.2016.01.010. PMID: 26780545.

22. Craven LJ, Nair Parvathy S, Tat-Ko J, Burton JP, Silverman MS. 2017; Extended screening costs associated with selecting donors for Fecal Microbiota Transplantation for treatment of metabolic syndrome-associated diseases. Open Forum Infect Dis. 4:ofx243. DOI: 10.1093/ofid/ofx243. PMID: 29255739. PMCID: PMC5730934.

23. Zhang X, Qiao J, Wu X, Da R, Zhao L, Wei Z. 2012; In vitro culture of Blastocystis hominis in three liquid media and its usefulness in the diagnosis of blastocystosis. Int J Infect Dis. 16:e23–8. DOI: 10.1016/j.ijid.2011.09.012. PMID: 22047715.