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DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick.
Acta Trop. 2019 Dec; 200:105185.AT

Abstract

Paragonimus westermani (P. westermani) is widely spread in Asian countries and is one of the most important causative agents for lung fluke diseases. The prevention and control of Paragonimiaisis mainly depends on the accurate diagnosis and effective treatment. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay targeted to a portion of the Ty3/gypsy-like LTR retrotransposon (Rn1) sequence coupled with a lateral flow dipstick (LFD) for the rapid detection of P. westermani-specific amplicons. The positive LAMP products were biotin-labeled and hybridized with a fluorescein isothiocyanate-labeled probe which could be visually detected by LFD. No cross-reaction were observed with other parasitic pathogens including Trichinella spiralis, Anisakis simplex, Schistosoma japonicum and Gnathostoma spinigerum, but this LAMP assay could not distinguish P. westermani with Paragonimus skrjabini and Paragonimus heterotremus. The detection limit of the LAMP assay for P. westermani was 2.7 fg/µL, while that of PCR method was 27 fg/µL. LAMP method was applied to detect P. westermani genomic DNA in blood samples form experimental infected dogs, and results showed the parasite was detectable as early as week 2. LAMP-LFD assay applicability was successfully tested in dog blood samples collected from five cities (Wenzhou, Hangzhou, Huzhou, Jiaxing and Shaoxing) in Zhejiang province. In summary, the established LAMP-LFD assay targeted to the Rn1 sequence is a rapid and convenient method for specific detection of P. westermani.

Authors+Show Affiliations

Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Anhui Institute of Schistosomiasis, Hefei, 230061, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China.Anhui Institute of Schistosomiasis, Hefei, 230061, China.Department of Immunity and Biochemistry, Institute of Parasitic Disease, Zhejiang Academy of Medical Sciences, Hangzhou, 310013, China. Electronic address: llsshh2003@163.com.

Pub Type(s)

Case Reports
Journal Article

Language

eng

PubMed ID

31542373

Citation

Xunhui, Zhuo, et al. "DNA Detection of Paragonimus Westermani: Diagnostic Validity of a New Assay Based On Loop-mediated Isothermal Amplification (LAMP) Combined With a Lateral Flow Dipstick." Acta Tropica, vol. 200, 2019, p. 105185.
Xunhui Z, Qingming K, Qunbo T, et al. DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick. Acta Trop. 2019;200:105185.
Xunhui, Z., Qingming, K., Qunbo, T., Haojie, D., Lesheng, Z., Di, L., Jianzu, D., Bin, Z., Rui, C., Tianping, W., & Shaohong, L. (2019). DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick. Acta Tropica, 200, 105185. https://doi.org/10.1016/j.actatropica.2019.105185
Xunhui Z, et al. DNA Detection of Paragonimus Westermani: Diagnostic Validity of a New Assay Based On Loop-mediated Isothermal Amplification (LAMP) Combined With a Lateral Flow Dipstick. Acta Trop. 2019;200:105185. PubMed PMID: 31542373.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - DNA detection of Paragonimus westermani: Diagnostic validity of a new assay based on loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick. AU - Xunhui,Zhuo, AU - Qingming,Kong, AU - Qunbo,Tong, AU - Haojie,Ding, AU - Lesheng,Zhang, AU - Di,Lou, AU - Jianzu,Ding, AU - Bin,Zheng, AU - Rui,Chen, AU - Tianping,Wang, AU - Shaohong,Lu, Y1 - 2019/09/19/ PY - 2018/12/03/received PY - 2019/09/11/revised PY - 2019/09/18/accepted PY - 2019/9/23/pubmed PY - 2020/1/29/medline PY - 2019/9/23/entrez KW - LAMP KW - LFD KW - Paragonimus westermani KW - Rn1 SP - 105185 EP - 105185 JF - Acta tropica JO - Acta Trop. VL - 200 N2 - Paragonimus westermani (P. westermani) is widely spread in Asian countries and is one of the most important causative agents for lung fluke diseases. The prevention and control of Paragonimiaisis mainly depends on the accurate diagnosis and effective treatment. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay targeted to a portion of the Ty3/gypsy-like LTR retrotransposon (Rn1) sequence coupled with a lateral flow dipstick (LFD) for the rapid detection of P. westermani-specific amplicons. The positive LAMP products were biotin-labeled and hybridized with a fluorescein isothiocyanate-labeled probe which could be visually detected by LFD. No cross-reaction were observed with other parasitic pathogens including Trichinella spiralis, Anisakis simplex, Schistosoma japonicum and Gnathostoma spinigerum, but this LAMP assay could not distinguish P. westermani with Paragonimus skrjabini and Paragonimus heterotremus. The detection limit of the LAMP assay for P. westermani was 2.7 fg/µL, while that of PCR method was 27 fg/µL. LAMP method was applied to detect P. westermani genomic DNA in blood samples form experimental infected dogs, and results showed the parasite was detectable as early as week 2. LAMP-LFD assay applicability was successfully tested in dog blood samples collected from five cities (Wenzhou, Hangzhou, Huzhou, Jiaxing and Shaoxing) in Zhejiang province. In summary, the established LAMP-LFD assay targeted to the Rn1 sequence is a rapid and convenient method for specific detection of P. westermani. SN - 1873-6254 UR - https://www.unboundmedicine.com/medline/citation/31542373/DNA_detection_of_Paragonimus_westermani:_Diagnostic_validity_of_a_new_assay_based_on_loop-mediated_isothermal_amplification_(LAMP)_combined_with_a_lateral_flow_dipstick L2 - https://linkinghub.elsevier.com/retrieve/pii/S0001-706X(18)31528-6 DB - PRIME DP - Unbound Medicine ER -