一种用于检测和区分溶组织内阿米巴、迪斯帕内阿米巴和莫氏内阿米巴的简单聚合酶链反应-限制性片段长度多态性技术的开发。

Development of a simple PCR-RFLP technique for detection and differentiation of E. histolytica, E. dispar and E. moshkovskii.

作者信息

Sardar Sanjib K, Kobayashi Seiki, Das Koushik, Saito-Nakano Yumiko, Dutta Shanta, Nozaki Tomoyoshi, Ganguly Sandipan

机构信息

Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India.

Department of Infectious Diseases, Keio University School of Medicine, Tokyo, Japan.

出版信息

Parasitol Res. 2023 Jan;122(1):139-144. doi: 10.1007/s00436-022-07706-1. Epub 2022 Oct 29.

DOI:10.1007/s00436-022-07706-1

PMID:36308533

Abstract

Epidemiological studies on amoebic infections are complicated by morphological overlap between the pathogenic E. histolytica, the commensal E. dispar and the amphizoic E. moshkovskii, necessitating molecular identification. The present study developed a simple and economical 18S PCR-RFLP method for the simultaneous detection and differentiation of the three species. PCR products were differentiated by Tat1 restriction digestion generating three different RFLP patterns. Validation was conducted by screening 382 faecal samples from human patients from Kolkata, India, hospitalized for diarrhoea. Analysis indicated that the PCR-RFLP could successfully differentiate between the three species and was confirmed by sequence analysis. This method could prove useful for clinical and epidemiological studies of amoebiasis.

摘要

由于致病性溶组织内阿米巴、共生性迪斯帕内阿米巴和兼性莫斯科维茨内阿米巴在形态上存在重叠，阿米巴感染的流行病学研究变得复杂，因此需要进行分子鉴定。本研究开发了一种简单且经济的18S PCR-RFLP方法，用于同时检测和区分这三种阿米巴。通过Tat1限制性消化对PCR产物进行区分，产生三种不同的RFLP模式。通过对来自印度加尔各答因腹泻住院的人类患者的382份粪便样本进行筛查来进行验证。分析表明，PCR-RFLP能够成功区分这三种阿米巴，并且通过序列分析得到了证实。该方法可能对阿米巴病的临床和流行病学研究有用。