Evaluation of immunoassays for the diagnosis of Schistosoma japonicum infection using archived sera (original) (raw)
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Parasites & Vectors, 2011
Background: The dipstick dye immunoassay (DDIA), recently commercially available in the People's Republic of China (P.R. China), is a rapid and simple test to detect human antibodies against Schistosoma Japonicum. Its performance and utility for screening schistosome infection in low endemic areas is little known. We therefore carried out a cross-sectional survey in seven villages with low endemicity of schistosomiasis in P.R. China and assessed the performance and utility of DDIA for diagnosis of schistosomiasis. Stool samples were collected and examined by the Kato-Katz method and the miracidium hatching technique. Serum samples, separated from whole blood of participants, were tested by DDIA.
Parasites & Vectors, 2011
Background: Schistosomiasis japonica (schistosomiasis) is a zoonosis that can seriously affect human health. At present, the immunodiagnostic assays for schistosomiasis detection are time-consuming and require well-trained personnel and special instruments, which can limit their use in the field. Thus, there is a pressing need for a simple and rapid immunoassay to screen patients on a large scale. In this study, we developed a novel rapid dipstick with latex immunochromatographic assay (DLIA) to detect anti-Schisaosoma japonicum antibodies in human serum. Results: Using latex microspheres as a color probe, DLIA was established to test standard positive and negative sera, in comparison with the classical enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of DLIA were 95.10% (97/102) and 94.91% (261/275), respectively. The cross-reaction rates with clonorchiosis, intestinal nematodes, Angiostrongylus cantonensis and paragonimiasis were 0, 0, 0 and 42.11% respectively. All the results showed no significant difference to the ELISA. In field tests, 333 human serum samples from an endemic area were tested with DLIA, and compared with ELISA and Kato-Katz method. There was no significant difference between DLIA and ELISA on positive and negative rates of detection; however, significant differences existed between DLIA and Kato-Katz method, and between ELISA and Kato-Katz method. The kappa value between DLIA and ELISA was 0.90.
American Journal of Tropical Medicine and Hygiene, 2009
More sensitive methods for diagnosing infection with Schistosoma japonicum are needed as control becomes more effective. We compared a real-time polymerase chain reaction (PCR) for stool samples with conventional diagnostic methods in a study of 1,727 persons from Anhui Province, China. Seroprevalence determined by using an indirect hemagglutination assay (IHA) was much higher (26.1%) than the prevalence in stool-based tests, which were 5.3%, 3.2%, and 3.0% for PCR, hatching test, and Kato-Katz thick smear, respectively. A large proportion of the positive stool samples were only positive in one or two tests. The PCR showed better agreement with IHA than the other two stool-based tests. A commonly used diagnostic algorithm with initial screening for antibodies and subsequent testing with the Kato-Katz thick smear of the seropositive results would have resulted in treatment of 22 people compared with 50 people if the PCR replaced the Kato-Katz thick smear. As prevalence and intensity decrease, the benefit of increased sensitivity using the PCR must be weighed against additional costs.
2021
The development of ELISA method to detect patients with schistosomiasis in Indonesia is a detection of the antigens that are derived from the peripheral vessels in the blood serum. This research is conducted in the Napu Plateau of Poso Regency, Central Sulawesi, Indonesia. The goal is to develop ELISA method with polyclonal antibodies to detect antigenic-excretory antigen of S. japonicum in patients with schistosomiasis. Activities in the study include activities in the field namely the isolation of worm S. japonicum and activities in laboratories such as production, characterization and purification by Bradford methods as well as electrophoresis test to see the protein Antigen profile. The results of the study of the 30 worm Schistosoma collected from 5 rats suffered schistosomiasis, obtained as much as 42.5 ml with a protein concentration of 1.351 mg/ml, and had two patterns of polypeptide tape with a molecular weight range of 20 and 39 Dalton kilo (kDa). The conclusion of the res...
The American Journal of Tropical Medicine and Hygiene, 2012
Sensitive Schistosoma japonicum detection methods are needed to progress from schistosomiasis control to elimination. The sensitivity of the Kato-Katz thick smear and miracidium hatching tests decrease with infection intensity and serological tests cannot always identify current infections. We evaluated a fecal polymerase chain reaction (PCR) assay to detect S. japonicum infection in 106 humans and 8 bovines in China. PCR was highly sensitive, detecting S. japonicum DNA at 0.5 eggs/g of stool. Comparing PCR examination of a single stool sample to the miracidium hatching test using three consecutive stool samples, more humans were hatching test positive (20%) than PCR positive (15%). However, two individuals were PCR positive in a village where no infections were detected by coprological methods. The sensitivity of PCR makes it a promising tool for schistosomiasis diagnostics and screening, although egg shedding variability and stool sample size present challenges for any detection method in low-transmission areas.
Real-time PCR diagnosis of Schistosoma japonicum in low transmission areas of China
Infectious diseases of poverty, 2018
Schistosomiasis in the People's Republic of China (PRC) can be traced back to antiquity. In the past 60 years, the Chinese government has made great efforts to control this persistent disease with elimination slated by 2020 through the implementation of a comprehensive control strategy. This strategy aims to reduce the role of bovines and humans as sources of infection as a pre-requisite for elimination through transmission interruption. The goal of elimination will be achievable only by the implementation of a sustainable surveillance and control system, with sensitive diagnosis a key feature so that the true disease burden is not underestimated. Currently used diagnostics lack the necessary sensitivity to accurately determine the prevalence of Schistosoma japonicum infection in areas with low infection intensities. It is of critical importance to find and treat people and to identify animals with low-level infections if the National Control Programme for China is to achieve sc...
PLoS ONE, 2012
Background: Schistosomiasis japonica remains a real threat to public health in China. The currently used immunodiagnostic assays are sensitive and have a certain degree of specificity, however, they all use complex crude antigens, are based on detection of schistosome-specific antibodies, and have been shown to cross-react with other parasitic diseases. Therefore, these assays cannot be used to evaluate chemotherapy efficacy. The development of highly sensitive and highly specific immunodiagnostic techniques that can monitor the decline of antibodies specific for S. japonica will be extremely valuable as part of the ongoing strategy to control schistosomiasis in endemic areas. Here we report on the identification of unique fraction antigens of soluble egg antigen (SEA) to which the antibodies disappear 7 weeks after effective treatment. Furthermore, we use these SEA fractions to develop a modified assay with both high sensitivity and specificity. Methodology/Principal Findings: SEA of S. japonicum was fractionated by electrophoresis using 7.5% SDS-PAGE under non-reducing conditions. The SEA fraction antigens to which antibodies were decreased soon after treatment were collected and used as the detection antigens to establish the FA-ELISA. Sera from patients with acute and chronic schistosomiasis infection, healthy people, and those with other parasitic diseases, were used to evaluate their sensitivity and specificity. Furthermore, sera from patients with chronic schistosomiasis infection were evaluated before and after treatment at different time points to evaluate their chemotherapeutic efficacy. Conclusion/Significance: We demonstrated that this novel FA-ELISA provided high sensitivity and specificity, with very low cross-reactivity, and can serve as an effective tool to determine the efficacy of chemotherapy against S. japonicum.
2014
Background Schistosomiasis remains a highly prevalent and serious parasitic disease. A major factor preventing its effective management is the scarcity of effective diagnostic tools. We did a genome-wide identification of diagnostic protein markers for schistosome infection and assessed their diagnostic validity in a field study. Methods We predicted putative secreted proteins of Schistosoma japonicum (SjSPs) and expressed them as glutathione S-transferase (GST)-fusion proteins. The fusion proteins were arrayed on glutathione (GSH)-immobilised microplates and screened with serum samples from patients with schistosomiasis diagnosed by the Kato-Katz method. We further assessed an identified protein marker for sensitivity and specificity, first in infected serum samples collected from Jiangxi and Hunan Provinces, China, and then through a field study, done in two villages located in a high schistosomiasis-endemic area of the southeast of China. Findings Of 204 recombinant proteins, 35 yielded seropositive reactions, eight showed strong immunoreactivity, and only one (SjSP-13) reacted to the entire panel of 14 archived samples. The reactivity of SjSP-13 to 476 serum samples showed 90·4% (95% CI 86·5–93·5) sensitivity and 98·9% (95% CI 95·9–99·9) specificity. Of 1371 residents enrolled in a field study from Dec 6, 2010, to June 23, 2011, only 74 individuals were identified as being egg-positive, whereas 465 were diagnosed as positive by the SjSP-13-based ELISA kit (rSP13-ELISA). Of the 394 individuals found egg-negative but rSP13-ELISA-positive, 363 (92·4%) were confirmed to be positive for schistosome infection by PCR detection of S japonicum SjR2 retrotransposon. Interpretation The application of this sensitive, specific, and affordable rSP13-ELISA method should help reduce schistosomiasis transmission through targeted treatment of individuals, particularly with low intensity infections, and therefore support schistosomiasis control and elimination strategies. Funding National 973 project in China.
Parasitology, 2009
SUMMARYPooled sample testing (PST) as a strategy for avoiding testing the majority of individual negative samples has been proposed for screening of diseases in low prevalence areas. There has been no standard guideline for PST in screening of Schistosoma japonicum infection of Yunnan, China. To document the optimum pool size with acceptable sensitivity of PST for screening of Schistosoma japonicum infection in this setting, an experimental pooling of each of 31 positive sera by IHA with various numbers of 24 negative sera was done. The results were used to create a statistical model which was subsequently used for simulation to predict sensitivity of the pooled serum tests in the population with varying prevalence and pool size. We found that to keep the sensitivity of PST above 90%, 1:05 should be the maximum dilution, that is, the optimum pool size should not be greater than 6. Antigen will have rather little interference if the prevalence of infection is low e.g. 1% or the antig...