Rapid Detection of Norovirus GII by Fluorescent Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) and Nanomagnetic Bead Separation

被引:0
作者
Li, Zhengkang [1 ]
Di, Yuwei [1 ]
Song, Xiaoyan [2 ]
Wu, Yanqi [3 ,4 ,5 ]
Feng, Yingye [6 ]
Zhang, Xinqiang [1 ]
Gong, Caiping [1 ]
Li, Guanghua [1 ]
机构
[1] Southern Med Univ, Guangdong Prov Peoples Hosp, Guangdong Acad Med Sci, Dept Clin Lab Med, Guangzhou 510080, Guangdong, Peoples R China
[2] Guangdong Prov Fertil Hosp, Guangdong Prov Reprod Sci Inst, NHC Key Lab Male Reprod & Genet, Guangzhou 510600, Guangdong, Peoples R China
[3] Hunan Univ Technol, Hunan Key Lab Biomed Nanomat & Devices, Zhuzhou 412007, Hunan, Peoples R China
[4] Macau Univ Sci & Technol, State Key Lab Qual Res Chinese Med, Taipa 999078, Macau, Peoples R China
[5] Shenzhen LemnisCare Med Technol Co Ltd, Shenzhen 518055, Guangdong, Peoples R China
[6] Guangzhou Med Univ, Guangdong Women & Childrens Hosp, Guangzhou 511400, Guangdong, Peoples R China
关键词
Norovirus; RT-LAMP; RT-PCR; PCR ASSAYS; PERFORMANCE;
D O I
10.1166/jbn.2023.3552
中图分类号
TB3 [工程材料学];
学科分类号
0805 ; 080502 ;
摘要
Noroviruses (NoVs) is the main cause of gastroenteritis in humans worldwide, mainly affecting school-age children and Copyright: American Scientific Publishers adults. NoVs are transmitted through feces and vomitus, includihuman contact, food, and water. Presently, NoVs Delivered by Ing enta are detected using molecular biological methods. Loop-mediated isothermal amplification (LAMP), specifically, requires little detection equipment, a short detection time, and low technical skills. Here, we established our own NoV reverse transcription (RT) polymerase chain reaction (PCR) quantitative detection system and a NoV GII RT-LAMP detection system. We collected 40 clinical samples, extracted RNAs, and used RT-PCR and RT-LAMP to detect NoV GII. The qualitative results of RT-LAMP were consistent with those of RT-PCR. However, a significant difference was observed between RT-LAMP and RT-PCR quantitative detection results. The NoV GII RT-LAMP detection system showed good sensitivity, up to 10(1), as well as good specificity. Furthermore, GI and GII did not interfere with each other. No false -positive responses were obtained for other gastrointestinal RNA viruses, such as Coxsackie virus A16 or enterovirus 71. Our results showed that the RT-LAMP detection system for NoV GII is suitable for the quantitative determination of NoV.
引用
收藏
页码:1413 / 1421
页数:9
相关论文
共 29 条
[1]   Coating Silica Layer on Fe3O4 Magnetic Nanoparticles and Application in Extracting High Quality Nucleic Acids from Blood Sample [J].
Ali, Zeeshan ;
Zou, Jun ;
Liu, Xiaolong ;
Bai, Yongkai ;
Hussain, Mubashir ;
Zhang, Lijun ;
Chen, Zhu ;
Chen, Hui ;
Li, Song ;
Deng, Yan ;
Zhang, Yuanying ;
Tang, Yongjun .
JOURNAL OF BIOMEDICAL NANOTECHNOLOGY, 2022, 18 (03) :828-836
[2]   Norovirus in Bottled Water Associated with Gastroenteritis Outbreak, Spain, 2016 [J].
Blanco, Albert ;
Guix, Susana ;
Fuster, Noemi ;
Fuentes, Cristina ;
Bartolome, Rosa ;
Cornejo, Thais ;
Maria Pinto, Rosa ;
Bosch, Albert .
EMERGING INFECTIOUS DISEASES, 2017, 23 (09) :1531-1534
[3]   Development and evaluation of a thermostatic nucleic acid testing device based on magnesium pyrophosphate precipitation for detecting Enterocytozoon hepatopenaei [J].
Chen, Zhu ;
Zhao, Kaixuan ;
He, Ziyu ;
Luo, Xiaofang ;
Qin, Zuodong ;
Tan, Yimin ;
Zheng, Xiangming ;
Wu, Zuozhong ;
Deng, Yan ;
Chen, Hui ;
Guo, Yuan ;
Li, Song .
CHINESE CHEMICAL LETTERS, 2022, 33 (08) :4053-4056
[4]  
China National Accreditati on Service for Conformity Assessment, 2018, CNAS-CL02-A009, P1
[5]  
Clinical and Laboratory Standards Institute, 2015, CLSI Document EP15-A2
[6]   Performance of Two Real-Time RT-PCR Assays for the Quantification of GI and GII Noroviruses and Hepatitis A Virus in Environmental Water Samples [J].
De Keuckelaere, Ann ;
Stals, Ambroos ;
Baert, Leen ;
Uyttendaele, Mieke .
FOOD ANALYTICAL METHODS, 2013, 6 (04) :1016-1023
[7]   A Targeted Metagenomics Approach to Study the Diversity of Norovirus GII in Shellfish Implicated in Outbreaks [J].
Desdouits, Marion ;
Wacrenier, Candice ;
Ollivier, Joanna ;
Schaeffer, Julien ;
Le Guyader, Francoise S. .
VIRUSES-BASEL, 2020, 12 (09)
[8]   Fast and Accurate Control Strategy for Portable Nucleic Acid Detection (PNAD) System Based on Magnetic Nanoparticles [J].
Fang, Yile ;
Liu, Haoran ;
Wang, Yue ;
Su, Xiangyi ;
Jin, Lian ;
Wu, Yanqi ;
Deng, Yan ;
Li, Song ;
Chen, Zhu ;
Chen, Hui ;
He, Nongyue .
JOURNAL OF BIOMEDICAL NANOTECHNOLOGY, 2021, 17 (03) :407-415
[9]   Genomics Analyses of GIV and GVI Noroviruses Reveal the Distinct Clustering of Human and Animal Viruses [J].
Ford-Siltz, Lauren A. ;
Mullis, Lisa ;
Sanad, Yasser M. ;
Tohma, Kentaro ;
Lepore, Cara J. ;
Azevedo, Marli ;
Parra, Gabriel I. .
VIRUSES-BASEL, 2019, 11 (03)
[10]   Performance of a one-step quantitative duplex RT-PCR for detection of rotavirus A and noroviruses GII during two periods of high viral circulation [J].
Fumian, Tulio M. ;
Leite, Jose Paulo G. ;
Rocha, Monica S. ;
de Andrade, Juliana S. R. ;
Fioretti, Julia M. ;
de Assis, Rosane M. S. ;
Assis, Matheus R. S. ;
Fialho, Alexandre M. ;
Miagostovich, Marize P. .
JOURNAL OF VIROLOGICAL METHODS, 2016, 228 :123-129