Colorimetric Detection of Dengue by Single Tube Reverse-Transcription-Loop-Mediated Isothermal Amplification

被引：61

作者：

Lau, Yee-Ling ^{[1
]}

Lai, Meng-Yee ^{[1
]}

Teoh, Boon-Teong ^{[2
]}

Abd-Jamil, Juraina ^{[2
]}

Johari, Jefree ^{[2
]}

Sam, Sing-Sin ^{[2
]}

Tan, Kim-Kee ^{[2
]}

AbuBakar, Sazaly ^{[2
]}

机构：

[1] Univ Malaya, Fac Med, Dept Parasitol, Trop Infect Dis Res & Educ Ctr, Kuala Lumpur 50603, Malaysia

[2] Univ Malaya, Fac Med, Dept Med Microbiol, Trop Infect Dis Res & Educ Ctr, Kuala Lumpur 50603, Malaysia

来源：

PLOS ONE | 2015年 / 10卷 / 09期

关键词：

WEST NILE VIRUS; RAPID DIAGNOSIS; DIFFERENTIATION; SEROTYPES; LAMP;

D O I：

10.1371/journal.pone.0138694

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Dengue is usually diagnosed by isolation of the virus, serology or molecular diagnostic methods. Several commercial kits for the diagnosis of dengue are existing, but concerns have arisen regarding to the affordability and performance characteristics of these kits. Hence, the loop-mediated isothermal amplification (LAMP) is potentially ideal to be used especially in resource limited environments. Serum was collected from healthy donors and patients diagnosed with dengue infection. RNA extracted from the serum samples were tested by reverse-transcription-LAMP assay developed based on 30-NCR gene sequences for DENV 1-4. Results were interpreted by a turbidity meter in real time or visually at the end of the assay. Sensitivity and specificity of RT-LAMP results were calculated and compared to qRT-PCR and ELISA. RT-LAMP is highly sensitive with the detection limit of 10 RNA copies for all serotypes. Dengue virus RNA was detected in all positive samples using RT-LAMP and none of the negative samples within 30-45 minutes. With continuing efforts in the optimization of this assay, RT-LAMP may provide a simple and reliable test for detecting DENV in areas where dengue is prevalent.

引用

页数：9

共 20 条

[1] Emergence of dengue virus type 4 genotype IIA in Malaysia [J].