Digital PCR: a new technology for diagnosis of parasitic infections

Pomari, Elena; Piubelli, Chiara; Perandin, Francesca; Bisoffi, Zeno

doi:10.1016/j.cmi.2019.06.009

Cited by 63 publications

(48 citation statements)

References 49 publications

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“…And cdPCR limits detection of HCMV viral load is 15 copies/ml in clinical sample. cdPCR complete PCR reaction through 2D array of microchamber, and cdPCR can realize three-color multiplexing ampli cation [25,26]. Due to simpli ed steps, cdPCR effectively reduce risk of contamination.…”

Section: Discussionmentioning

confidence: 99%

The Prevalence of Human Cytomegalovirus Infection in Children Leukemia by Chip Digital PCR

Wang

Mao

et al. 2021

Preprint

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Background To establish a method for detecting HCMV viral load to guide clinical treatment by chip digital PCR (cdPCR). Methods 5.67×106TCID50/ml of HCMV AD169 was serially diluted to evaluate sensitive of cdPCR, HSV-1, HSV-2, VZV, EBV, HHV-6 and HHV-7 were used to evaluate the specificity of HCMV cdPCR. HCMV infection were analyzed in 110 children leukemia whole blood by RT-qPCR and cdPCR. Results The sensitive of HCMV cdPCR was up to 71 ± 32 copies/ml, which is higher than that of RT-qPCR. HCMV cdPCR did not cross react with other herpesviruses. The cdPCR effectively detected 7 HCMV positive samples, making the laboratory diagnosis rate of HCMV increased by 6.36% (7/110) for children leukemia patients. And the prevalence of HCMV infection increased from 28.18–34.54% in 110 children leukemia patients by cdPCR. Conclusion cdPCR is more sensitive to detect viral load than RT-qPCR. The cdPCR may be used to evaluate relationship between viral load and progression of HCMV infection in patients.

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Section: Discussionmentioning

confidence: 99%

The Prevalence of Human Cytomegalovirus Infection in Children Leukemia by Chip Digital PCR

Wang

Mao

et al. 2021

Preprint

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“…The limitations of microscopy and antigen detection tests have encouraged parasitologists to move towards the use of genomic amplification methods made possible with the advent of molecular diagnostics, but they still remain underused 2 , 11 . Step by step, antigen detection tests for G. lamblia , Cryptosporidium spp and Entamoeba hystolitica have been widely introduced into the day-to-day laboratory workflow, especially in those laboratories with lower capacities for parasitological diagnosis.…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, there has been a tremendous effort to focus research on the development of new diagnostic methods, such as serological, molecular, and proteomic approaches 1 , but these techniques also have limitations due to the fact that they do not detect all possible parasites; also they are expensive and are not available for all clinical microbiology laboratories. Molecular assays have emerged as the solution for diagnosis of a lot of infectious diseases and several targets have been used for the diagnosis of parasitic infections but, many of them are locally designed solutions and they are not standardized 2 .…”

Section: Introductionmentioning

confidence: 99%

Automated parasitological diagnosis in clinical microbiology laboratories

Fernández‐Rivas

Rivaya

Romaní

et al. 2021

Sci Rep

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Although there is a low prevalence of parasitological infections in Europe, the diagnosis of intestinal parasites is still difficult and laborious for microbiology laboratories. Currently, antigen detection assays and molecular biology allow a more accurate diagnosis, but these techniques have limitations as they cannot detect all the possible parasites present in the samples. The objective of the study was to evaluate the accuracy and the usefulness of automated microscopy SediMAX2 (77 Elektronika, Budapest, Hungary) in the detection of parasitic infections from feces. A total of 197 formol-fixed stool samples were processed in parallel by wet mount examination and by SediMAX2. Sensitivities, specificities and predictive values were analyzed, reaching a sensitivity of 89.51% and a specificity of 98.15% and a very good positive predictive value (99.22%). SediMAX2 is a good tool for a reliable diagnosis of intestinal parasitic infections. The rapid processing and the flexibilty of storage of images analyzed make its incorporation into the day to day laboratory routine recommendable.

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“…In the case of patients with very low parasitaemia, a method that can improve the sensitivity of the molecular tests is desirable. Recent evidence has demonstrated that ddPCR may be especially useful for the screening of low-level parasitaemia due to its analytical sensitivity [18] compared especially to quantitative rtPCR. It has been shown that quantitative rtPCR is more sensitive than thick-film microscopy in whole blood and has a good agreement also with plasma samples [24].…”

Section: Discussionmentioning

confidence: 99%

“…Target genes include 18S rRNA (the most frequently used), tRNA, cytochrome b, ama1, and stevor [17]. On the basis of recent advances in molecular biology and keeping in mind the limitations of conventional methods, additional techniques have been introduced for the DNA-based detection of Plasmodium, such as loop-mediated isothermal amplification (LAMP) and droplet digital PCR (ddPCR) [18]. Studies have shown the efficacy of all these molecular techniques in detecting Plasmodium DNA extracted from whole blood.…”

Section: Introductionmentioning

confidence: 99%

Droplet Digital PCR for the Detection of Plasmodium falciparum DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods

et al. 2020

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Background: The estimation of Plasmodium falciparum 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 Plasmodium, 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 P. falciparum 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, p < 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 P. falciparum DNA, with 77% positive samples among malaria subjects. Conclusion: Absolute quantitation by ddPCR can be a flexible technique for Plasmodium detection, with potential application in the diagnosis of malaria. In particular, ddPCR is a powerful approach for Plasmodium DNA analysis on serum when blood samples are unavailable.

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Digital PCR: a new technology for diagnosis of parasitic infections

Cited by 63 publications

References 49 publications

The Prevalence of Human Cytomegalovirus Infection in Children Leukemia by Chip Digital PCR

The Prevalence of Human Cytomegalovirus Infection in Children Leukemia by Chip Digital PCR

Automated parasitological diagnosis in clinical microbiology laboratories

Droplet Digital PCR for the Detection of Plasmodium falciparum DNA in Whole Blood and Serum: A Comparative Analysis with Other Molecular Methods

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