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Malaria prevalence defined by microscopy, antigen detection, DNA amplification and total nucleic acid amplification in a malaria-endemic region during the peak malaria transmission season

机译:疟疾流行高峰期通过显微镜,抗原检测,DNA扩增和总核酸扩增确定的疟疾流行率

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摘要

objectives To determine the malaria prevalence by microscopy, antigen detection and nucleic acid detection in a defined subpopulation in a Plasmodium falciparum-endemic region during the peak transmission season. methods Blood specimens were collected in a cross-sectional study involving children aged 5-10 years (n = 195) presenting with acute fever to two clinics in Western Kenya. All specimens underwent microscopy, HRP2 and aldolase antigen detection by enzyme immunoassay (EIA), parasite-specific DNA and total nucleic acid (RNA and DNA) by real-time PCR (qPCR) and reverse-transcriptase PCR (qRT-PCR). results Microscopy detected 65/195 cases of malaria infection [95% confidence interval (CI) 52-78]. HRP2 and aldolase EIA had similar sensitivity levels detecting antigen in 65/195 (95% CI, 52-78) and 57/195 (95% CI, 45-70) cases. Discordants in antigen detection vs. microscopy occurred at < 470 parasites/mu1 and <4900 parasites/mul for HRP2 and aldolase, respectively. Detection of total nucleic acid allowed a 3 log lower limit of detection than just DNA detection by real-time PCR in vitro. In clinical specimens, 114/195 (95% CI, 100-127) were qPCR positive (DNA), and 187/195 (95% CI, 179-191) were qRT-PCR positive (DNA plus RNA). conclusions The prevalence of submicroscopic malaria infection was significantly higher when detecting total nucleic acid than just DNA in this outpatient population during the high transmission season. Defining standards for submicroscopic infection will be important for control programmes, diagnostics development efforts and molecular epidemiology studies.
机译:目的通过在传播高峰期的恶性疟原虫流行区的特定亚群中,通过显微镜检查,抗原检测和核酸检测来确定疟疾患病率。方法在一项横断面研究中收集了血液样本,该研究涉及肯尼亚西部两个诊所的5-10岁(195例)急性发烧儿童。所有样品均进行了显微镜检查,HRP2和醛缩酶抗原通过酶免疫测定(EIA),寄生虫特异性DNA和总核酸(RNA和DNA)的实时PCR(qPCR)和逆转录酶PCR(qRT-PCR)检测。结果显微镜检出65/195例疟疾感染[95%置信区间(CI)52-78]。 HRP2和醛缩酶EIA在65/195(95%CI,52-78)和57/195(95%CI,45-70)病例中检测抗原的敏感性水平相似。 HRP2和醛缩酶分别在<470寄生虫/μl和<4900寄生虫/μl时发生抗原检测与显微镜检查不一致。总核酸的检测比体外通过实时PCR的DNA检测所允许的检测限低3个对数。在临床标本中,qPCR阳性(DNA)为114/195(95%CI,100-127),qRT-PCR阳性(DNA加RNA)为187/195(95%CI,179-191)。结论在高传播季节期间,在该门诊人群中检测总核酸时,亚显微疟疾感染的发生率明显高于仅DNA。定义亚显微感染的标准对于控制程序,诊断开发工作和分子流行病学研究将非常重要。

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