Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods
Background: The estimation of <i>Plasmodium falciparum</i> parasitaemia can vary according to the method used. Recently, droplet digital PCR (ddPCR) has been proposed as a promising approach in the molecular quantitation of <i>Plasmodium</i>, but its ability to predict the ac...
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MDPI AG
2020-06-01
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| Online Access: | https://www.mdpi.com/2076-0817/9/6/478 |
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| author | Elena Pomari Ronaldo Silva Lucia Moro Giulia La Marca Francesca Perandin Federica Verra Zeno Bisoffi Chiara Piubelli |
| author_facet | Elena Pomari Ronaldo Silva Lucia Moro Giulia La Marca Francesca Perandin Federica Verra Zeno Bisoffi Chiara Piubelli |
| author_sort | Elena Pomari |
| collection | DOAJ |
| description | Background: The estimation of <i>Plasmodium falciparum</i> parasitaemia can vary according to the method used. Recently, droplet digital PCR (ddPCR) has been proposed as a promising approach in the molecular quantitation of <i>Plasmodium</i>, but its ability to predict the actual parasitaemia on clinical samples has not been largely investigated. Moreover, the possibility of applying the ddPCR-sensitive method to serum samples has never been explored. Methods: We used, for the first time, ddPCR on both blood and serum to detect the DNA of <i>P. falciparum</i> in 52 paired samples from 26 patients. ddPCR was compared with loop-mediated isothermal amplification (LAMP) and rtPCR. The correlation between the ddPCR results, microscopy, and clinical parameters was examined. Results: ddPCR and microscopy were found to be strongly correlated (ρ(26) = 0.83111, <i>p</i> < 0.0001) in blood. Samples deviating from the correlation were partially explained by clinical parameters. In serum samples, ddPCR revealed the best performance in detecting <i>P. falciparum</i> DNA, with 77% positive samples among malaria subjects. Conclusion: Absolute quantitation by ddPCR can be a flexible technique for <i>Plasmodium</i> detection, with potential application in the diagnosis of malaria. In particular, ddPCR is a powerful approach for <i>Plasmodium</i> DNA analysis on serum when blood samples are unavailable. |
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| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T19:07:10Z |
| publishDate | 2020-06-01 |
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| series | Pathogens |
| spelling | doaj.art-7a9507a540184d1daceded2b5b18e46f2023-11-20T04:05:56ZengMDPI AGPathogens2076-08172020-06-019647810.3390/pathogens9060478Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular MethodsElena Pomari0Ronaldo Silva1Lucia Moro2Giulia La Marca3Francesca Perandin4Federica Verra5Zeno Bisoffi6Chiara Piubelli7Department of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyDepartment of Infectious, Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Negrar di Valpolicella, 37024 Verona, ItalyBackground: The estimation of <i>Plasmodium falciparum</i> parasitaemia can vary according to the method used. Recently, droplet digital PCR (ddPCR) has been proposed as a promising approach in the molecular quantitation of <i>Plasmodium</i>, but its ability to predict the actual parasitaemia on clinical samples has not been largely investigated. Moreover, the possibility of applying the ddPCR-sensitive method to serum samples has never been explored. Methods: We used, for the first time, ddPCR on both blood and serum to detect the DNA of <i>P. falciparum</i> in 52 paired samples from 26 patients. ddPCR was compared with loop-mediated isothermal amplification (LAMP) and rtPCR. The correlation between the ddPCR results, microscopy, and clinical parameters was examined. Results: ddPCR and microscopy were found to be strongly correlated (ρ(26) = 0.83111, <i>p</i> < 0.0001) in blood. Samples deviating from the correlation were partially explained by clinical parameters. In serum samples, ddPCR revealed the best performance in detecting <i>P. falciparum</i> DNA, with 77% positive samples among malaria subjects. Conclusion: Absolute quantitation by ddPCR can be a flexible technique for <i>Plasmodium</i> detection, with potential application in the diagnosis of malaria. In particular, ddPCR is a powerful approach for <i>Plasmodium</i> DNA analysis on serum when blood samples are unavailable.https://www.mdpi.com/2076-0817/9/6/478<i>Plasmodium falciparum</i>DNAabsolute quantitationdroplet digital PCRmicroscopy |
| spellingShingle | Elena Pomari Ronaldo Silva Lucia Moro Giulia La Marca Francesca Perandin Federica Verra Zeno Bisoffi Chiara Piubelli Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods Pathogens <i>Plasmodium falciparum</i> DNA absolute quantitation droplet digital PCR microscopy |
| title | Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods |
| title_full | Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods |
| title_fullStr | Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods |
| title_full_unstemmed | Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods |
| title_short | Droplet Digital PCR for the Detection of <i>Plasmodium falciparum</i> DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods |
| title_sort | droplet digital pcr for the detection of i plasmodium falciparum i dna in whole blood and serum a comparative analysis with other molecular methods |
| topic | <i>Plasmodium falciparum</i> DNA absolute quantitation droplet digital PCR microscopy |
| url | https://www.mdpi.com/2076-0817/9/6/478 |
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