Highly sensitive detection of malaria parasitemia in a malaria-endemic setting: performance of a new loop-mediated isothermal amplification kit in a remote clinic in Uganda.
Highly sensitive detection of malaria parasitemia in a malaria-endemic setting: performance of a new loop-mediated isothermal amplification kit in a remote clinic in Uganda.
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在疟疾流行环境中对疟疾寄生虫血症的高度敏感性检测：在乌干达偏远诊所中新的环路介导的等温扩增试剂盒的性能。
DOI：
10.1093/infdis/jit184
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发表时间：
2013-08-15
期刊：
影响因子：
--
通讯作者：
Bell D
中科院分区：
文献类型：
--
作者：
Hopkins H;González IJ;Polley SD;Angutoko P;Ategeka J;Asiimwe C;Agaba B;Kyabayinze DJ;Sutherland CJ;Perkins MD;Bell D
背景：目前的疟疾诊断测试，包括显微镜和抗原检测快速测试，不能可靠地检测低密度感染。 聚合酶链反应等分子方法非常敏感，但对于实地部署来说仍然过于复杂。一种新的商业分子检测的基础上环介导的等温扩增（LAMP）进行了评估，现场使用。方法：对来自乌干达乡村诊所的272名门诊患者的样本进行疟疾LAMP（日本Eiken Chemical）评价，并与专家显微镜、巢式PCR和定量PCR（qPCR）进行比较。 两名技术人员在3天培训后使用2种替代血液样品制备方法进行测定，并通过荧光测定法对结果进行目视判读。结果：与3孔巢式PCR相比，LAMP和单孔巢式PCR的敏感性均为90%，显微镜检查的敏感性为51%。 对于恶性疟原虫qPCR滴度≥2个寄生虫/µL的样本，LAMP灵敏度为97.8%（95%置信区间，93.7%-99.5%）。大多数假阴性LAMP结果涉及寄生虫血症水平可通过3孔巢式PCR检测但通过qPCR检测非常低或不可检测的样品。结论：疟疾LAMP在乌干达一个偏远的诊所达到了与英国参考实验室的单孔巢式PCR相似的灵敏度。 LAMP大大降低了在疟疾流行环境中可达到的检测阈值，为消除战略中的诊断、监测和筛查提供了新的工具。
Background. Current malaria diagnostic tests, including microscopy and antigen-detecting rapid tests, cannot reliably detect low-density infections. Molecular methods such as polymerase chain reaction (PCR) are highly sensitive but remain too complex for field deployment. A new commercial molecular assay based on loop-mediated isothermal amplification (LAMP) was assessed for field use. Methods. Malaria LAMP (Eiken Chemical, Japan) was evaluated for samples from 272 outpatients at a rural Ugandan clinic and compared with expert microscopy, nested PCR, and quantitative PCR (qPCR). Two technicians performed the assay after 3 days of training, using 2 alternative blood sample–preparation methods and visual interpretation of results by fluorescence assay. Results. Compared with 3-well nested PCR, the sensitivity of both LAMP and single-well nested PCR was 90%; the microscopy sensitivity was 51%. For samples with a Plasmodium falciparum qPCR titer of ≥2 parasites/µL, LAMP sensitivity was 97.8% (95% confidence interval, 93.7%–99.5%). Most false-negative LAMP results involved samples with parasitemia levels detectable by 3-well nested PCR but very low or undetectable by qPCR. Conclusions. Malaria LAMP in a remote Ugandan clinic achieved sensitivity similar to that of single-well nested PCR in a United Kingdom reference laboratory. LAMP dramatically lowers the detection threshold achievable in malaria-endemic settings, providing a new tool for diagnosis, surveillance, and screening in elimination strategies.
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影响因子：
3
作者：
Manjurano A;Okell L;Lukindo T;Reyburn H;Olomi R;Roper C;Clark TG;Joseph S;Riley EM;Drakeley C
通讯作者：
Drakeley C
影响因子：
3.7
作者：
Lucchi NW;Demas A;Narayanan J;Sumari D;Kabanywanyi A;Kachur SP;Barnwell JW;Udhayakumar V
通讯作者：
Udhayakumar V
DOI：
10.4269/ajtmh.2011.10-0507
发表时间：
2011-06-01
影响因子：
3.3
作者：
Campos, Ivon M.;Uribe, Mary L.;Maestre, Amanda
通讯作者：
Maestre, Amanda
影响因子：
3.7
作者：
Ouedraogo, Andre Lin;Bousema, Teun;Sauerwein, Robert
通讯作者：
Sauerwein, Robert
影响因子：
6.4
作者：
Hopkins, Heidi;Bebell, Lisa;Dorsey, Grant
通讯作者：
Dorsey, Grant