Prospective European-wide multicentre study on a blood based real-time PCR for the diagnosis of acute schistosomiasis

Wichmann, Dominic; Poppert, Sven; Thien, Heidrun von; Clerinx, Joannes; Dieckmann, Sebastian; Jensenius, Mogens; Parola, Philippe; Richter, Joachim; Schunk, Mirjam; Stich, August; Zanger, Philipp; Burchard, Gerd; Tannich, Egbert

doi:10.1186/1471-2334-13-55

Cited by 73 publications

(60 citation statements)

References 23 publications

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“…5,[11][12][13][14] In this study, we are assessing the effect of concentration by centrifugation and frozen storage of urine samples on the sensitivity of a Schistosoma real-time PCR assay routinely used in our laboratory.…”

mentioning

confidence: 99%

Impact of Short-Time Urine Freezing on the Sensitivity of an Established Schistosoma Real-Time PCR Assay

Kenguele¹,

Adegnika²,

Nkoma³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. Urogenital schistosomiaisis is a serious public health problem in sub-Saharan Africa. In this study, we have updated an established real-time polymerase chain reaction (PCR) routinely used in our laboratory. Schistosoma genusspecific real-time PCR was performed on DNA isolated from 85 urine samples and pellets obtained after centrifugation without and after frozen storage. The results revealed that concentration by centrifugation of the urine samples and freezing of the samples before extracting DNA improves the sensitivity of the PCR.

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mentioning

confidence: 99%

Impact of Short-Time Urine Freezing on the Sensitivity of an Established Schistosoma Real-Time PCR Assay

Kenguele¹,

Adegnika²,

Nkoma³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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“…In Schistosoma spp. infections, positive qPCR responses to blood products are generally achieved during the acute phase of infections (Wichmann et al, 2013;Zhou et al, 2011) where the initial host immune response leads to degradation of parasite stages and higher levels of circulating DNA.…”

Section: Discussionmentioning

confidence: 99%

“…No parasite DNA could be detected in limited stored blood samples. While real-time PCR has been successfully used in the diagnosis of other parasite infections in human blood samples, including the systemic coccidian Toxoplasma gondii (Ricciardi and Ndao, 2015;RobertGangneux and Dardé, 2012) and blood flukes Schistosoma spp (Cavalcanti et al, 2013;Wichmann et al, 2013), these methods are generally most effective in the acute phase of infections (Wichmann et al, 2013;Zhou et al, 2011) where increased host immune response to a new insult results in parasite degradation and increased circulating parasite DNA. Given the high prevalence of Neospirorchis spp.…”

Section: Aim 4 -Explore and Develop Potential Methods For The Diagnosmentioning

confidence: 99%

“…Traditionally, the focus has been on faecal examination and antigen based tests, however, these have limited application in posttreatment monitoring, as egg shedding and serologic markers often persist following successful treatment. PCR-based approaches aimed at detection of DNA in faeces and blood have been shown to offer higher sensitivity and specificity than conventional faecal egg detection in stool (Lier et al, 2008;Oliveira et al, 2010;Pontes et al, 2002;Pontes et al, 2003) and serological approaches (Pontes et al, 2002;Wichmann et al, 2009;Wichmann et al, 2013). PCR may be particularly useful in areas of low endemicity or where low burdens or levels of egg excretion are present (Gentile et al, 2011;Oliveira et al, 2010;Pontes et al, 2002) and also in early stages of disease (Wichmann et al, 2013).…”

Section: Development Of Techniques For Sea Turtle Parasite Investigationmentioning

confidence: 99%

“…(Cavalcanti et al, 2013;Gentile et al, 2011;Lier et al, 2008;Wang et al, 2011;Wichmann et al, 2013). Real-time PCR utilising…”

Section: Development Of Techniques For Sea Turtle Parasite Investigationmentioning

confidence: 99%

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Diversity, impacts and diagnosis of pathogenic parasites in sea turtles from Queensland, Australia

Chapman¹

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Parasitism is a significant cause of stranding and mortality in endangered sea turtles. Two parasites in particular (or groups thereof) have been noted for their impacts on green sea turtle populations. The first are the spirorchiid blood flukes, which inhabit the circulatory systems of their host. The second is the eimeriid coccidian Caryospora cheloniae, a systemic parasite affecting a number of organs. Diversity among these organisms is poorly explored, and is almost certainly greater than currently recognised. The parasites and their associated diseases fluctuate both temporally and spatially in terms of prevalence and severity, however, the factors driving these fluctuations are poorly understood. A variety of questions remain unanswered in terms of the epidemiology and relative pathogenicity of the different parasite species. Of particular interest are brain lesions, which have been associated with both parasites and in some cases lead to neurological deficiency and subsequent mortality. Difficulties in finding answers to these questions is compounded by a lack of reliable, fast and quantitative ante-and post-mortem diagnostic tools for either parasite, which restricts their investigation.In order to catalogue local species and assess diversity, adult spirorchiid flukes were collected during post-mortem examination of deceased sea turtles and characterised by molecular and morphological means. Eleven distinct species or genotypes were identified, comprising a mix of previously described and novel species or variants. Samples were also collected from green sea turtles that died during a coccidiosis related mass mortality event in south east Queensland and northern New South Wales. While only one species of coccidian (C. cheloniae) had been previously described, molecular characterisation of the organism implicated in the outbreak revealed two distinct coccidian genotypes, which has significant implications for diagnosis and management.Spirorchiid ova have been associated with granulomatous lesions in a wide range of host tissues. However, it has not been possible to identify spirorchiids beyond the genus level based on morphology, which has frustrated any attempts to identify the species or variants responsible for severe lesions. In this thesis, a molecular approach was used and a terminal restriction fragment length polymorphism (T-RFLP) assay was developed to detect and identify individual species within the often mixed assemblages of ova in turtle tissues. This assay proved to be a more specific and sensitive alternative to traditional iii microscopic detection methods. Through correlation with histopathology and gross pathology, the tissue tropisms, relative occurrence and pathogenicity of each species were investigated. The most common species (Neospirorchis Genotype 2) was found in 96% of samples, encompassing tissues from all organs sampled. On average, a greater number of spirorchiid species were detected in tissues where lesions were present, and numbers increased along with the severity o...

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Diagnosis of Human Trematode Infections

Esteban

Muñoz-Antolí

Toledo

et al. 2014

Advances in Experimental Medicine and Biology

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Prospective European-wide multicentre study on a blood based real-time PCR for the diagnosis of acute schistosomiasis

Cited by 73 publications

References 23 publications

Impact of Short-Time Urine Freezing on the Sensitivity of an Established Schistosoma Real-Time PCR Assay

Impact of Short-Time Urine Freezing on the Sensitivity of an Established Schistosoma Real-Time PCR Assay

Diversity, impacts and diagnosis of pathogenic parasites in sea turtles from Queensland, Australia

Diagnosis of Human Trematode Infections

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