Detection and pathological role of intestinal protozoa in children

Intestinal parasites are considered a growing public health problem, being protozoa the main cause of intestinal disease. The objective of our study is to compare the detection of intestinal protozoa by microscopy versus real-time PCR, as well as to determine the most prevalent protozoa in our environment in the paediatric population.

An observational longitudinal study was carried out, both by microscopy and real time-PCR in stool samples from children (0- 15 years) received from April 2019 to March 2021.Children were classified in two groups according if they had or not had clinical parasitosis. Microscopic examination was performed in all samples using the Ritchie concentration technique with the commercial Mini PARASEP system (Movaco-Grifols®). The presence of Cryptosporidium sp. was evaluated with the modified Ziehl-Neelsen acid-fast stain. The real-time PCR was performed to all samples using the Allplex ™ gastrointestinal parasite panel 4 (Seegene®).

During the study period, 500 samples were received, being positive 31 (6.2%) by microscopy and 256 (51.2 %) by PCR. By microscopy, Blastocystis hominis was the most frequently observed (4.8%), followed by Giardia lamblia (1.6%), Dientamoeba fragilis (0.2%) and Cryptosporidium species (0.2%). Regarding the identification by PCR, D. fragilis (35.2%) was mainly identified, followed by B. hominis (28.1%), G. lamblia (7%) and Cryptosporidium sp. (0.8%) without finding clear differences in aetiology according to age. In the case of B. hominis and D. fragilis, there were not differences in the detection of these protozoa between the control group and children with clinical parasitosis (p = 0.11).

Real-time PCR increases the detection of intestinal protozoa, being underdiagnosed by microscopy, especially D. fragilis, in which PCR is considered the most appropriate method for its detection.