PCR-Based Detection of Enterocytozoon bieneusi in Diarrheic Patients from Guangdong, Shandong, Shanghai, and Zhejiang Provinces, China: A Study on Prevalence and Genotypic Characteristics

Jiang-qiong Ke¹Sun Lijie²Qi Yu³Xiaorong Yu⁴Zhongkai Zhang¹Aiying Jiang¹Xin Peng¹Jiabin Zhang¹Fuhao Jiang¹Yanyan Jiang^5*Huanhuan Zhou^4*Wei Zhao^1*

• ¹Wenzhou Medical University, Wenzhou, China
• ²The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China
• ³Shanghai Pudong New Area People's Hospital, Shanghai, China
• ⁴Laboratory Department, Qingdao South District Center for Disease Control and Prevention, Qingdao, China
• ⁵Chinese Center for Disease Control and Prevention National Institute of Parasitic Diseases, Shanghai, China

Introduction: Enterocytozoon bieneusi is the most prevalent microsporidian species infecting humans and causing diarrhea. Epidemiological investigations seldom focus on this pathogen, and its disease burden has been underestimated. This research investigated the prevalence and genotypic characteristics of E. bieneusi in patients with diarrhea from Guangdong (Zhuhai), Shandong (Qingdao), Shanghai and Zhejiang (Wenzhou) Provinces, China. Methods: A total of 691 fecal specimens from patients exhibiting diarrhea were collected and subjected to polymerase chain reaction (PCR) detection, targeting the internal transcribed spacer (ITS) region of the E. bieneusi genome. Genotypes were identified by sequencing PCR products, and zoonotic risk was evaluated through homology and phylogenetic analysis. Results: 4.9% (34/691) of patients were positive for E. bieneusi. The patients from Shanghai had the highest incidence at 14.3% (21/147), which was significantly higher than those in Qingdao (2.9%, 5/171), Wenzhou (3.3%, 3/90), and Zhuhai (1.8%, 5/283) (χ2 = 35.5, P < 0.001). Meanwhile, the positive rate was 7.1% (18/254), 3.1% (10/325) and 5.4% (6/112) among children, adults, and the elderly, respectively. Additionally, the positive rate of patients in rural areas was 8.6% (26/302), significantly higher than that in urban areas, which was 2.1% (8/389) (χ2 = 15.6, P < 0.001). Twelve genotypes of E. bieneusi were identified, including seven known genotypes: CHG3 (n = 17), CHG19 (n = 3), EbPigITS7 (n = 3), TypeIV (n = 3), CHG5 (n = 1), EbpA (n = 1), and S7 (n = 1), as well as five novel genotypes (SHH1, WZH1, ZHH1, ZHH2 and ZHH3), each represented by a single sample. The identified genotypes can be categorized into Groups 1, 2, 12, and 13 through phylogenetic analysis. Conclusions: This study offers insights into the epidemiology and genetic diversity of E. bieneusi among patients experiencing diarrhea in four provinces of China. It also underscores the necessity of ongoing monitoring and management to prevent the spread of this pathogen.