Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

被引:132
作者
Domingo, Cristina [1 ]
Patel, Pranav [1 ]
Yillah, Jasmin [1 ]
Weidmann, Manfred [2 ]
Mendez, Jairo A. [3 ]
Nakoune, Emmanuel Rivalyn [4 ]
Niedrig, Matthias [1 ]
机构
[1] Robert Koch Inst, Berlin, Germany
[2] Univ Med Ctr, Dept Virol, Gottingen, Germany
[3] Inst Nacl Salud, Virol Lab, Bogota, Colombia
[4] Inst Pasteur, Bangui, Cent Afr Republ
关键词
INFECTION; ASSAY;
D O I
10.1128/JCM.01799-12
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Reported methods for the detection of the yellow fever viral genome are beset by limitations in sensitivity, specificity, strain detection spectra, and suitability to laboratories with simple infrastructure in areas of endemicity. We describe the development of two different approaches affording sensitive and specific detection of the yellow fever genome: a real-time reverse transcription-quantitative PCR (RT-qPCR) and an isothermal protocol employing the same primer-probe set but based on helicase-dependent amplification technology (RT-tHDA). Both assays were evaluated using yellow fever cell culture supernatants as well as spiked and clinical samples. We demonstrate reliable detection by both assays of different strains of yellow fever virus with improved sensitivity and specificity. The RT-qPCR assay is a powerful tool for reference or diagnostic laboratories with real-time PCR capability, while the isothermal RT-tHDA assay represents a useful alternative to earlier amplification techniques for the molecular diagnosis of yellow fever by field or point-of-care laboratories.
引用
收藏
页码:4054 / 4060
页数:7
相关论文
共 23 条
[21]  
Vainio J., 1998, YELLOW FEVER
[22]   Helicase-dependent isothermal DNA amplification [J].
Myriam Vincent ;
Yan Xu ;
Huimin Kong .
The EMBO Reports, 2004, 5 (8) :795-800
[23]   Improved LNA probe-based assay for the detection of African and South American yellow fever virus strains [J].
Weidmann, Manfred ;
Faye, Ousmane ;
Faye, Oumar ;
Kranaster, Ramon ;
Marx, Andreas ;
Nunes, Marcio R. T. ;
Vasconcelos, Pedro F. C. ;
Hufert, Frank T. ;
Sall, Amadou A. .
JOURNAL OF CLINICAL VIROLOGY, 2010, 48 (03) :187-192