Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle

Agneessens, Joost; Claerebout, Edwin; Vercruysse, Jozef

doi:10.1016/s0304-4017(01)00398-3

Cited by 14 publications

(16 citation statements)

References 25 publications

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“…Unfortunately, these immunological methods also have their limitations. Parasite ES products are released in the GI tract and these antigens may be digested during its passage (Agneessens et al 2001). Another problem with these methods is the cross-reactivity with other concurring GI nematodes (Johnson et al 1996) or other parasites (Agneessens et al 2001).…”

Section: Discussionmentioning

confidence: 99%

“…Copro-antigen capture enzyme-linked immunosorbent assays take into account current intestinal infection with adult worms because these methods detect excretory/secretory (ES) antigens released by the metabolic active stages (Agneessens et al 2001;Deplazes et al 1992;Roepstorff 1998). Unfortunately, these immunological methods also have their limitations.…”

Section: Discussionmentioning

confidence: 99%

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Which McMaster egg counting technique is the most reliable?

et al. 2011

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This study focuses on the comparison of three selected modifications of the McMaster counting technique, namely the McMaster method modified by Wetzel (W) and Zajíček (Z), as well as the concentration McMaster technique according to Roepstorff and Nansen (R&N). These modifications differ in the weights of faeces examined (W, 2 g/Z, 1 g/R&N, 4 g), flotation solutions (W, NaCl/Z, MgSO(4) + Na(2)S(2)O(3)/R&N, NaCl + glucose), centrifugation (W, none/Z, 2,000 RPM for 2 min and 2,000 RPM for 1 min/R&N, 1,200 RPM for 5 min), number of McMaster chambers investigated (W, 3/Z, 2/R&N, 2), and multiplication factors used (W, 67/Z, 33/R&N, 20). To investigate the sensitivity and reliability of these methods, nematode eggs (Teladorsagia circumcincta) were used. Parasite elements are distributed through negative binomial distribution in naturally infected host faeces, and the number of parasite elements in a given amount of faeces sample is unknown to man. Therefore, we decided to prepare the exact number of eggs which were added to the parasite negative faeces; the faecal sample was then investigated. From this perspective, this is the first time a comparison of the McMaster methods has been so accurately investigated. This approach allows us to evaluate the real sensitivity and reliability of the tested method. As the findings of this study indicate, the highest sensitivity and reliability were obtained using the Roepstorff and Nansen modification. This McMaster modification is able to detect 20 eggs per sample (in 70% of samples). Concentrations of 200 and 500 eggs can be found in almost 100% of samples. Moreover, this method is simple, cheap and fast. For these reasons, we can recommend this method for routine veterinary practice.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Which McMaster egg counting technique is the most reliable?

et al. 2011

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“…Accordingly, if simplified and inexpensive protocols are developed, the immunologic detection of hookworm coproantigens may represent an attractive diagnostic alternative to fecal egg assays. Coproantigen ELISAs have previously been reported for nematodes such as Haemonchus contortus 25 and Teladorsagia circumcincta 26 in sheep, Ostertagia ostertagi in cattle, 27 Heligmosomoides polygyrus 28 and Trichinella spiralis 29 in mice, and Strongyloides ratti in rats. 30 However, to our knowledge this is the first report of the technique being applied to the diagnosis and evaluation of experimental hookworm infection.…”

Section: Discussionmentioning

confidence: 99%

“…Asterisks below the brackets indicate level of significance: *P < 0.01 or **P < 0.001. nematode species. [26][27][28]30 Although additional studies of the sensitivity and specificity of the fecal ELISA will be necessary if the assay is to be adapted for use as a human diagnostic test, these preliminary results suggest that the fecal ELISA may eventually offer a means to diagnose low intensity infections that might not be detected by conventional microscopy, allowing for more accurate estimates of hookworm prevalence. We noted that OD values were generally lower when ES was diluted in fecal extract as opposed to PBS-T, indicating some inhibition of the assay by fecal material; similar findings have been reported for the previously cited fecal ELISAs for other nematodes.…”

Section: Discussionmentioning

confidence: 99%

Detection of Excretory/Secretory Coproantigens in Experimental Hookworm Infection

Bungiro

Cappello

2005

The American Journal of Tropical Medicine and Hygiene

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This report describes the detection of hookworm excretory/secretory (ES) antigens in soluble hamster fecal extracts by an enzyme-linked immunosorbent assay (ELISA). A rabbit polyclonal IgG antibody against Ancylostoma ceylanicum ES was used to capture hookworm coproantigens that were then detected using pooled, high-titer, infected hamster serum. The ELISA was capable of detecting ES proteins over a range of 10 ng/mL to 10 mug/mL when the antigens were diluted in buffer or uninfected fecal extract, and ES could be detected in infected hamster feces at dilutions up to 1:256. Examination of the kinetics of coproantigen production demonstrated that detectable amounts of ES were produced as early as four days after A. ceylanicum infection, whereas fecal eggs were not observed until day 17. Moreover, fecal ES levels correlated well with intestinal worm burden and could be detected in wet or dry stool samples stored for 14 days over a temperature range of -80 degrees C to 37 degrees C. The fecal ELISA was then adapted to analyze the excretion of AceES-2, a novel immunogenic ES protein recently cloned from A. ceylanicum cDNA. AceES-2 was found to be excreted in feces with kinetics similar to that of whole ES. Examination of individual hookworm antigens by this method will provide new insights into the molecular host-parasite interaction and may form the basis for future diagnostic methods.

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“…There have also been reports where assay sensitivity and/or specificity was unsatisfactory e.g. detection of Strongyloies ratti [23], Haemonchus contortus [24] and Ostertagia ostertagi [25]. With the exception of the O. Ostertagia assay, assays of satisfactory performance have employed specific antiserum raised against excretory/secretory (E/S) antigens of the respective parasites, rather than utilizing antiserum raised against total somatic antigen.…”

Section: Introductionmentioning

confidence: 99%

A Coproantigen Diagnostic Test for Strongyloides Infection

Sykes

McCarthy

2011

PLoS Negl Trop Dis

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Accurate diagnosis of infection with the parasite Strongyloides stercoralis is hampered by the low concentration of larvae in stool, rendering parasitological diagnosis insensitive. Even if the more sensitive agar plate culture method is used repeated stool sampling is necessary to achieve satisfactory sensitivity. In this manuscript we describe the development of a coproantigen ELISA for diagnosis of infection. Polyclonal rabbit antiserum was raised against Strongyloides ratti excretory/secretory (E/S) antigen and utilized to develop an antigen capture ELISA. The assay enabled detection of subpatent rodent S. ratti and human S. stercoralis infection. No cross-reactivity was observed with purified E/S from Schistosoma japonicum, the hookworms Ancylostoma caninum, A. ceylanicum, nor with fecal samples collected from rodents harboring Trichuris muris or S. mansoni infection. Strongyloides coproantigens that appear stable when frozen as formalin-extracted fecal supernatants stored at −20°C remained positive up to 270 days of storage, whereas supernatants stored at 4°C tested negative. These results indicate that diagnosis of human strongyloidiasis by detection of coproantigen is an approach worthy of further development.

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Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle

Cited by 14 publications

References 25 publications

Which McMaster egg counting technique is the most reliable?

Which McMaster egg counting technique is the most reliable?

Detection of Excretory/Secretory Coproantigens in Experimental Hookworm Infection

A Coproantigen Diagnostic Test for Strongyloides Infection

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