Jiang Yanyan, Ren Jinhua, Yuan Zhongying, Liu Aiqin, Zhao Hong, Liu Hua, Chu Lei, Pan Wei, Cao Jianping, Lin Yijin, Shen Yujuan
BMC Infect Dis. 2014 Oct 25;14:555. doi: 10.1186/s12879-014-0555-7.
Cryptosporidium hominis and C. parvum are usually considered to be the major pathogens responsible for human cryptosporidiosis. However, there have been few studies regarding the molecular epidemiology of Cryptosporidium in human infections in China. Here we investigated Cryptosporidium infection in patients with diarrhea, in Danyang Hospital of Jiangsu Province, China, at the genotype level.
A total of 232 stool specimens were collected from outpatients with diarrhea in Danyang Hospital of Jiangsu Province, China, from February 2012 to January 2013. Each specimen was stained from direct fecal smears and examined for Cryptosporidium using modified acid fast staining and microscopy. Moreover, genomic DNA of each fecal sample was screened for the presence of Cryptosporidium with nested PCR, which was genotyped by analyzing the DNA sequences of small subunit rRNA (SSU rRNA).
The average infection rate of Cryptosporidium was 1.3% (3/232) by microscopy and subjected to PCR amplification of the SSU rRNA gene of Cryptosporidium, with 9.91% (23/232) being positive for Cryptosporidium with a significant peak in autumn. Based on the SSU rRNA gene, two Cryptosporidium spp. were identified, including C. andersoni (n =21) and C. hominis (n =2). Two types of C. andersoni, designated as A370 + and A370 - , were found in the SSU rRNA gene in our present study, which was 100% homologous to C. andersoni infections derived from dairy calves and goats, respectively. The clinical questionnaires showed no significant difference in age, gender and frequency of diarrhea, but duration of diarrhea was shorter for C. andersoni than that of C. hominis (mean, 2 vs. 4 days; p <0.01).
C. andersoni is the dominant species in Danyang City of Jiangsu Province. The fact that SSU rRNA sequences of C. andersoni obtained from human stools exhibited 100% homologous to those derived from dairy calves and goats supported that C. andersoni infection might be attributable to animal origin. The difference in the duration of diarrhea of C. andersoni and C. hominis indicated that different Cryptosporidium species might cause different clinical manifestations.
人隐孢子虫和微小隐孢子虫通常被认为是导致人类隐孢子虫病的主要病原体。然而,关于中国人类感染隐孢子虫的分子流行病学研究较少。在此,我们在基因水平上调查了中国江苏省丹阳市医院腹泻患者中的隐孢子虫感染情况。
2012年2月至2013年1月期间,从中国江苏省丹阳市医院的腹泻门诊患者中收集了232份粪便标本。每份标本均制成直接粪便涂片,采用改良抗酸染色和显微镜检查隐孢子虫。此外,使用巢式PCR检测每份粪便样本的基因组DNA中是否存在隐孢子虫,并通过分析小亚基核糖体RNA(SSU rRNA)的DNA序列进行基因分型。
显微镜检查隐孢子虫的平均感染率为1.3%(3/232),对隐孢子虫的SSU rRNA基因进行PCR扩增,隐孢子虫阳性率为9.91%(23/232),秋季出现显著高峰。基于SSU rRNA基因,鉴定出两种隐孢子虫,包括安氏隐孢子虫(n = 21)和人隐孢子虫(n = 2)。在本研究中,在SSU rRNA基因中发现了两种类型的安氏隐孢子虫,分别命名为A370 +和A370 - ,它们分别与来自奶牛犊和山羊的安氏隐孢子虫感染100%同源。临床问卷调查显示,在年龄、性别和腹泻频率方面无显著差异,但安氏隐孢子虫感染患者的腹泻持续时间比人隐孢子虫感染患者短(平均,2天对4天;p < 0.01)。
安氏隐孢子虫是江苏省丹阳市的优势种。从人类粪便中获得的安氏隐孢子虫的SSU rRNA序列与来自奶牛犊和山羊的序列100%同源,这一事实支持安氏隐孢子虫感染可能源于动物。安氏隐孢子虫和人隐孢子虫腹泻持续时间的差异表明,不同的隐孢子虫物种可能导致不同的临床表现。