Comparison of next-generation droplet digital PCR (ddPCR) with quantitative PCR (qPCR) for enumeration of Cryptosporidium oocysts in faecal samples

Yang, Rongchang; Paparini, Andrea; Monis, Paul; Ryan, Una

doi:10.1016/j.ijpara.2014.08.004

Cited by 162 publications

(143 citation statements)

References 41 publications

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“…An expansion on qPCR techniques is multiplex PCR, which amplifies more than one DNA target in a single reaction mixture and has been used successfully for case detection (197), for differentiating between S. japonicum, S. mansoni, and S. haematobium, and as an important tool in epidemiological studies and in monitoring schistosomiasis control programs (197,206). In a recent advance in PCR technology, droplet digital PCR (dd PCR) was developed and has proven to be more sensitive and precise than qPCR (211)(212)(213). dd PCR has been used successfully for the detection of cellfree DNA and in the diagnosis of infections and other clinical conditions (211,214,215).…”

Section: Detection Of Schistosoma Dna By Pcrmentioning

confidence: 99%

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Advances in the Diagnosis of Human Schistosomiasis

et al. 2015

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SUMMARY Schistosomiasis is a major neglected tropical disease that afflicts more than 240 million people, including many children and young adults, in the tropics and subtropics. The disease is characterized by chronic infections with significant residual morbidity and is of considerable public health importance, with substantial socioeconomic impacts on impoverished communities. Morbidity reduction and eventual elimination through integrated intervention measures are the focuses of current schistosomiasis control programs. Precise diagnosis of schistosome infections, in both mammalian and snail intermediate hosts, will play a pivotal role in achieving these goals. Nevertheless, despite extensive efforts over several decades, the search for sensitive and specific diagnostics for schistosomiasis is ongoing. Here we review the area, paying attention to earlier approaches but emphasizing recent developments in the search for new diagnostics for schistosomiasis with practical applications in the research laboratory, the clinic, and the field. Careful and rigorous validation of these assays and their cost-effectiveness will be needed, however, prior to their adoption in support of policy decisions for national public health programs aimed at the control and elimination of schistosomiasis.

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Section: Detection Of Schistosoma Dna By Pcrmentioning

confidence: 99%

“…In a recent advance in PCR technology, droplet digital PCR (dd PCR) was developed and has proven to be more sensitive and precise than qPCR (211)(212)(213). dd PCR has been used successfully for the detection of cellfree DNA and in the diagnosis of infections and other clinical conditions (211,214,215).…”

Section: Detection Of Schistosoma Dna By Pcrmentioning

confidence: 99%

Advances in the Diagnosis of Human Schistosomiasis

et al. 2015

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“…All samples were screened using an 18S qPCR as previously described (Yang et al, 2014) and a C. parvum and C. hominis specific qPCR at a unique Cryptosporidium specific gene (Clec) coding for a novel mucin-like glycoprotein that contains a Ctype lectin domain (CTLD) previously described (Morgan et al, 1996;Bhalchandra et al, 2013;Yang et al, 2009;. This was done to determine if there were any mixed infections with C. parvum and/or C. hominis in the samples.…”

Section: Molecular Typingmentioning

confidence: 99%

Genetic characterization of Cryptosporidium in animal and human isolates from Jordan

Hijjawi

Mukbel

Yang

et al. 2016

Veterinary Parasitology

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(Mukbel R). #Joint first authors Highlights First genotyping study of Cryptosporidium in animals in Jordan  Faecal samples (n=284) from domestic animals and humans (n=48) screened  Overall prevalence of 11.6% in animals and six species detected.  Novel Cryptosporidium genotype detected in horses AbstractLittle is known about the epidemiology of Cryptosporidium in Jordan and to date, only one genotyping study has been conducted on Cryptosporidium isolates from Jordanian children. In the present study, a total of 284 faecal samples from Jordanian cattle, sheep, goats and chicken and 48 human faecal samples were screened for the presence of Cryptosporidium using an 18S quantitative PCR (qPCR) and a C. parvum/C. hominis specific qPCR at a lectin locus. Of these, 37 of 284 animal faecal samples were positive by qPCR at the 18S locus giving an overall prevalence of 11.6%. The point prevalence of Cryptosporidium in chickens, sheep, horses, cattle and goats ranged from 4.8% (chickens) to 18.7% (cattle). A total of six species were detected; C. xiaoi (n=9), C.andersoni (n=7), C. ryanae (n=5), C. parvum (n=4), C. baileyi (n=1) and a genetically distinct and potentially novel species in two isolates from horses. Sub-genotype analysis of the 4 C. parvum isolates at the 60-kDa glycoprotein (gp60) locus identified subtype IIaA19G2R1 (n=2) and IIaA16GR1 (n=2). For the human samples, 4 positives (8.3% prevalence) were detected. Of these, two were C. parvum (subtypes IIdA20G1 and IIaA15G2R1) and two were C. hominis (subtypes 1bA9G3 and 1bA10G2R2). Further studies are required to better understand the epidemiology and transmission of Cryptosporidium in Jordan.

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“…The mean RSQ (R squared) was 0.99 and the % Relative Standard Deviation (RDS) = 1.5% (Yang et al, 2014a). The Cryptosporidium actin qPCR described was also evaluated using haemocytometer counted oocysts and also flow-cytometry counted oocysts on clinical cryptosporidiosis samples and again found to be both precise and reliable (Yang et al, 2014c).…”

Section: Pcr Amplification -Cryptosporidium and Giardiamentioning

confidence: 94%

Greater intensity and frequency of Cryptosporidium and Giardia oocyst shedding beyond the neonatal period is associated with reductions in growth, carcase weight and dressing efficiency in sheep

Jacobson

Williams

Yang

et al. 2016

Veterinary Parasitology

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Haemonchus status (positive or negative) with lamb production were assessed using general linear models (HSCW and dressing %) and linear mixed effects models (live weight). HighCryptosporidium parvum shedding was associated with lower live weight, ranging 2.31-4.52kg over the 3 sampling occasions. Cryptosporidium parvum shedding was associated 3 with less HSCW in high (3.22kg less) and low (3.22kg less) shedding lambs post-weaning, and high (2.21kg less) and low (2.60kg less) shedding lambs pre-slaughter as well as lower dressing % (2.7% lower in high shedding lambs post-weaning). Cryptosporidium (all species) shedding pre-slaughter was associated with reduced dressing % in both high (1.25% lower) and low (1.21% lower) shedding lambs. Giardia shedding pre-slaughter was associated with 0.59kg less HSCW in high shedding lambs. Increased frequency of C.parvum and Giardia shedding in a specific animal (repeated detection) were associated with Abbreviations:HSCW: hot standard carcase weight WEC: Faecal worm egg count

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Comparison of next-generation droplet digital PCR (ddPCR) with quantitative PCR (qPCR) for enumeration of Cryptosporidium oocysts in faecal samples

Cited by 162 publications

References 41 publications

Advances in the Diagnosis of Human Schistosomiasis

Advances in the Diagnosis of Human Schistosomiasis

Genetic characterization of Cryptosporidium in animal and human isolates from Jordan

Greater intensity and frequency of Cryptosporidium and Giardia oocyst shedding beyond the neonatal period is associated with reductions in growth, carcase weight and dressing efficiency in sheep

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