Vascular wilt, caused by Verticillium dahliae and V. longisporum, limits the quality and yield of agricultural crops. Although quantitative real-time PCR (qPCR) has greatly improved the diagnosis of these two pathogens over traditional, time-consuming isolation methods, the relatively poor detection sensitivity and high measurement bias for traceable matrix-rich samples need to be improved. Here, we thus developed a droplet digital PCR (ddPCR) assay for accurate, sensitive detection and quantification of V. dahliae and V. longisporum. We compared the analytical and diagnostic performance in detail of ddPCR and the corresponding qPCR assay against the genomic DNA (gDNA) of the two fungi from cultures and field samples. In our study, the species specificity, quantification linearity, analytical sensitivity, and measurement viability of the two methods were analyzed. The results indicated that ddPCR using field samples enhanced diagnostic sensitivity, decreased quantification bias, and indicated less susceptibility to inhibitors compared with qPCR. Although ddPCR was as sensitive as qPCR when using gDNA from cultures of V. dahliae and V. longisporum, its detection rates using field samples were much higher than those of qPCR, potentially due to the inhibition from residual matrix in the extracts. The results showed that digital PCR is more sensitive and accurate than qPCR for quantifying trace amounts of V. dahliae and V. longisporum and can facilitate management practices to limit or prevent their prevalence.
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
Xu, Xiaoli
Peng, Cheng
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
Peng, Cheng
Wang, Xiaofu
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
Wang, Xiaofu
Chen, Xiaoyun
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
Chen, Xiaoyun
Wang, Qiang
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
Wang, Qiang
Xu, Junfeng
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Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R ChinaZhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, Hangzhou 310021, Zhejiang, Peoples R China
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CNRS UMR 5089, F-31077 Toulouse, France
Inst Pharmacol & Biol Struct, F-31077 Toulouse, France
Univ Toulouse, F-31077 Toulouse, France
UPS, F-31077 Toulouse, France
IPBS, F-31077 Toulouse, FranceCNRS UMR 5089, F-31077 Toulouse, France
Braikia, Fatima-Zohra
Chemin, Guillaume
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CNRS UMR 5089, F-31077 Toulouse, France
Inst Pharmacol & Biol Struct, F-31077 Toulouse, France
Univ Toulouse, F-31077 Toulouse, France
UPS, F-31077 Toulouse, France
IPBS, F-31077 Toulouse, FranceCNRS UMR 5089, F-31077 Toulouse, France
Chemin, Guillaume
Moutahir, Mohammed
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CNRS UMR 5089, F-31077 Toulouse, France
Inst Pharmacol & Biol Struct, F-31077 Toulouse, France
Univ Toulouse, F-31077 Toulouse, France
UPS, F-31077 Toulouse, France
IPBS, F-31077 Toulouse, FranceCNRS UMR 5089, F-31077 Toulouse, France
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Shandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Wang, Cong
Ding, Qiang
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Ding, Qiang
Plant, Pamela
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Plant, Pamela
Basheer, Mayada
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Univ Toronto, Dept Lab Med & Pathobiol, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Basheer, Mayada
Yang, Chuance
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Yang, Chuance
Tawedrous, Eriny
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Tawedrous, Eriny
Krizova, Adriana
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, Canada
Univ Toronto, Dept Lab Med & Pathobiol, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Krizova, Adriana
Boulos, Carl
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Boulos, Carl
Farag, Mina
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Farag, Mina
Cheng, Yufeng
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Shandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R ChinaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
Cheng, Yufeng
Yousef, George M.
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St Michaels Hosp, Keenan Biomed Res Ctr, Li Ka Shing Knowledge Inst, Toronto, ON, Canada
Univ Toronto, Dept Lab Med & Pathobiol, Toronto, ON, CanadaShandong Univ, Dept Radiat Oncol, Qilu Hosp, Jinan 250012, Shandong, Peoples R China
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Ewha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South KoreaEwha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South Korea
Kim, Tae Gwan
Jeong, So-Yeon
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Ewha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South KoreaEwha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South Korea
Jeong, So-Yeon
Cho, Kyung-Suk
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Ewha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South KoreaEwha Womans Univ, Dept Environm Sci & Engn, Global Res Program Top5, Seoul 120750, South Korea