Subtype analysis of Blastocystis isolates using SSU rRNA-DNA sequencing in rural and urban population in southern Turkey

Abstract

Blastocystis is a common and emerging parasite often seen in many studies conducted in urban population, with scanty reports on rural communities. However, little is known about the public health significance of Blastocystis infection. A total of 28 Blastocystis isolates from 17 (17/28, 60.71%) patients living in rural area and 11 (11/28, 39.29%) patients living in urban area were screened with seven kinds of sequenced-tagged site primers for identification of subtype. PCR products were sequenced with same combination of primers using the BigDye Terminator V 3.1 cycle sequencing kit, as per the manufacturers’ protocol on the 3730 DNA analyzer (Applied Biosystems, Carlsbad CA, USA). The cross-comparison of the Blastocystis sequences of samples were determined by the neighbor-joining method based on a distance matrix between sequence pairs to generate dendograms. The following subtypes were identified; subtype 1 (10/28, 35.7%), subtype 3 (7/28, 25.0%), subtype 2 (5/28, 17.8%), subtype 4 (3/28, 10.7%), subtype 5 (1/28, 3.6%), subtype 6 (1/28, 3.6%), and subtype 7 (1/28, 3.6%) in all DNA samples. The comparison of Blastocystis subtypes distribution among the patients from rural and urban area revealed subtype 5 (1/17, 5.9%), subtype 6 (1/17, 5.9%) and subtype 7 (1/17, 5.9%) from patients of rural area but not any of these subtypes in patients living urban area. This study is the first large-scale study to examine the occurrence of Blastocystis in Turkey to shed lights on the cosmopolitan distribution of Blastocystis subtypes in southern part of Turkey. Subtype 5, subtype 6 and subtype 7 were determined in only rural area. The findings of this study suggest that Blastocystis is transmitted from animal to human and possess a zoonotic potential.