Lacante S A, Jiang C, Mustamir A A, Mizuno T, Bi X, Syafruddin D, Tokoro M
Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
Graduate School of Hasanuddin University, Jl. Perintis Kemerdekaan Km.10, Makassar, Sulawesi Selatan, Indonesia.
MethodsX. 2024 Jul 23;13:102875. doi: 10.1016/j.mex.2024.102875. eCollection 2024 Dec.
, a human intestinal protozoan parasite of the diplomonad group, has been overlooked because of its commensal features; therefore, molecular studies on this parasite are limited. To address this gap, we designed a molecular screening protocol using polymerase chain reaction (PCR) and DNA sequencing targeting the 18S small subunit ribosomal RNA gene and applied this screening method to the molecular epidemiological analysis of spp. in humans and various livestock. We validated our methodology using stool samples collected from 215 humans and 270 animal hosts (buffaloes, pigs, dogs, goats, horses, rodents, chickens, and ducks) during an annual epidemiological investigation conducted from 2013 to 2016 on Sumba Island, Indonesia. The overall prevalences of spp. were 33.9 % ( 73/215) in humans and 25.2 % ( 68/270) in mammals and avians. The positive predictive value of this PCR method for spp., as evaluated through sequencing, was 90.1 % in human samples and 58.1 % in non-human samples (particularly low, 11.4 % in rodents). Although the specificity of the PCR approach may not be perfect, in combination with DNA sequencing, it was effective in detecting and identifying a partial sequence (1458 bp) of the target gene region in species.
作为双滴虫类的一种人体肠道原生动物寄生虫，由于其共生特性而被忽视；因此，对这种寄生虫的分子研究有限。为了填补这一空白，我们设计了一种分子筛查方案，使用聚合酶链反应（PCR）和针对18S小亚基核糖体RNA基因的DNA测序，并将这种筛查方法应用于人体和各种家畜中该物种的分子流行病学分析。在2013年至2016年于印度尼西亚松巴岛进行的年度流行病学调查期间，我们使用从215名人类和270只动物宿主（水牛、猪、狗、山羊、马、啮齿动物、鸡和鸭）收集的粪便样本验证了我们的方法。该物种的总体患病率在人类中为33.9%（73/215），在哺乳动物和鸟类中为25.2%（68/270）。通过测序评估，这种PCR方法对该物种的阳性预测值在人类样本中为90.1%，在非人类样本中为58.1%（在啮齿动物中特别低，为11.4%）。尽管PCR方法的特异性可能并不完美，但与DNA测序相结合，它有效地检测和鉴定了该物种目标基因区域的部分序列（1458 bp）。
