Genotypic and Phylogenetic Profile of Cryptosporidium Parvum Strains from HIV Positive Patients in Maiduguri, Nigeria

M. Y. Iliyasu, U. M. Askira, I. M. Tom, M. Sahal, S. Y. Dogonjeji, H. Tahir, Z. M. Kabeer, H. S. Musa, S. Panda, A. Sama’ila
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Abstract

Background: Cryptosporidium is an etiologic agent of gastro-intestinal disorders and severe, profuse watery diarrhoea in immunocompromised patients, such as people living with HIV/AIDS. Aim: This study was carried out to determine the genotypic and phylogenetic characteristics of Cryptosporidium parvum among HIV patients who usually received antiretroviral therapy (ART) in four selected hospitals in Maiduguri Metropolis. Study Design: This is a clinical laboratory study of opportunistic pathogens in HIV/AIDS Patients. Place and Duration of Study: This was conducted at the Department of Medical Laboratory Science, the University of Maiduguri in collaboration with the Biological Sciences Department, ATBU Bauchi, Nigeria, between March 2021, and February 2022. Methods: A total of 400 faecal specimens were collected from confirmed HIV-positive patients and screened for Cryptosporidium-specific antigen by Enzyme Link Immunosorbent Assay (ELISA). Genomic DNA Extraction was done by Quick-DNATM Faecal Microbe Miniprep with PCR amplification and sequencing of 18s SSU rRNA gene using specific reference primers. Results: Out of the 70 HIV/AIDS and Cryptosporidium co-infected patients, 20 of the samples contained 18s SSU rRNA genes, with the highest frequency (45.0%) found in patients from the University Teaching Hospital. The results of phylogenetic analyses indicated that there is significant intra-species diversity in the genus Cryptosporidium. The four human C. parvum isolates differ from the bovine and the two avian isolates in three regions of the 18s rRNA gene. Conclusion: The human genotype (genotype I) found in this study is exclusively human and in a single non-human primate, in bovine genotype (genotype II) has proved to be anthroponotic and zoonotic to the livestock. The study, therefore, advocates further genotyping of Cryptosporidium from both HIV-infected patients and immunocompetent hosts from various regions together with surveillance of animal and environmental reservoirs is highly recommended.
尼日利亚迈杜古里地区HIV阳性患者小隐孢子虫基因型及系统发育分析
背景:隐孢子虫是免疫功能低下患者(如艾滋病毒/艾滋病感染者)胃肠道疾病和严重、大量水样腹泻的病原。目的:研究在迈杜古里市选定的四家医院接受抗逆转录病毒治疗(ART)的HIV患者中细小隐孢子虫的基因型和系统发育特征。研究设计:这是一项HIV/AIDS患者机会致病菌的临床实验室研究。研究地点和时间:研究于2021年3月至2022年2月在迈杜古里大学医学实验室科学系与尼日利亚包奇大学生物科学系合作进行。方法:收集400例hiv确诊阳性患者粪便标本,采用酶联免疫吸附试验(ELISA)筛选隐孢子虫特异性抗原。基因组DNA提取采用Quick-DNATM粪便微生物Miniprep, PCR扩增,特异参考引物测序18s SSU rRNA基因。结果:在70例HIV/AIDS和隐孢子虫合并感染的患者中,有20例样本含有18s个SSU rRNA基因,其中以大学教学医院患者的频率最高(45.0%)。系统发育分析结果表明,隐孢子虫属具有明显的种内多样性。4个人分离的小孢子虫在18s rRNA基因的3个区域与牛分离的小孢子虫和2个禽分离的小孢子虫不同。结论:在本研究中发现的人类基因型(基因型I)完全是人类和单一的非人类灵长类动物,在牛基因型(基因型II)已被证明是人畜共患。因此,该研究提倡对来自不同地区的hiv感染患者和免疫能力强的宿主进行进一步的隐孢子虫基因分型,并强烈建议对动物和环境宿主进行监测。
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