Diagnosis Of Cystic Echinococcosis Research Articles

Immunohistochemistry as a Reliable Tool for the Diagnosis of Cystic Echinococcosis in Patients from Sardinia, Italy—A Confirmatory Study

Cystic Echinococcosis (CE) is a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.). This study aims to investigate the use of two monoclonal antibodies (mAbEmG3 and mAbEm2G11) by immunohistochemistry (IHC) to confirm the diagnosis of CE in human patients, in particular in those cases in which other techniques fail to provide a correct or conclusive diagnosis. For this purpose, a survey on 13 patients was performed. These subjects were referred to Sardinian hospitals (Italy) from 2017 to 2022 and were suspected to be affected by CE. Our findings from these 13 patients showed the detection of E. granulosus sensu stricto by IHC in 12 of 13 echinococcal cysts, as one sample was of a non-parasitological origin. The results confirmed that IHC, by means of the mAbEmG3 and mAbEm2G11, is a reliable diagnostic tool that showed a very high performances when tested on strain of E. granulosus s.l. from Sardinia.

Ultrasound diagnosis of cystic echinococcosis: updates and implications for clinical management.

The diagnosis of cystic echinococcosis (CE) is based on imaging. Detection of a focal lesion with morphological characteristics of Echinococcus granulosus sensu lato metacestode is the starting point for the diagnostic workup. In organs explorable with ultrasound (US), this is the method of choice for both aetiological diagnosis of CE and staging of the CE cyst. Staging in terms of lesion morphology is also needed when serology is added to the diagnostic workflow when imaging alone is inconclusive. Finally, staging guides the clinical management of uncomplicated CE, especially in the liver. This commentary provides an overview of the most up-to-date evidence backing the above-mentioned role of US in the diagnosis and clinical management of CE. Finally, we outline future perspectives for the improvement of CE diagnosis.

Comparison of Methods in the Serologic Diagnosis of Cystic Echinococcosis.

Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests. In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard. Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ2 = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test. There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability.

Rapid and non-invasive detection of cystic echinococcosis in sheep based on serum fluorescence spectrum combined with machine learning algorithms.

Cystic echinococcosis (CE) is a grievous zoonotic parasitic disease. Currently, the traditional technology of screening CE is laborious and expensive, developing an innovative technology is urgent. In this study, we combined serum fluorescence spectroscopy with machine learning algorithms to develop an innovative screening technique to diagnose CE in sheep. Serum fluorescence spectra of Echinococcus granulosus sensu stricto-infected group (n = 63) and uninfected E. granulosus s.s. group (n = 60) under excitation at 405 nm were recorded. The linear support vector machine (Linear SVM), Quadratic SVM, medium radial basis function (RBF) SVM, K-nearest neighbor (KNN), and principal component analysis-linear discriminant analysis (PCA-LDA) were used to analyze the spectra data. The results showed that Quadratic SVM had the great classification capacity, its sensitivity, specificity, and accuracy were 85.0%, 93.8%, and 88.9%, respectively. In short, serum fluorescence spectroscopy combined with Quadratic SVM algorithm has great potential in the innovative diagnosis of CE in sheep.

Machine learning-assisted serum SERS strategy for rapid and non-invasive screening of early cystic echinococcosis.

Early and accurate diagnosis of cystic echinococcosis (CE) with existing technologies is still challenging. Herein, we proposed a novel strategy based on the combination of label-free serum surface-enhanced Raman scattering (SERS) spectroscopy and machine learning for rapid and non-invasive diagnosis of early-stage CE. Specifically, by establishing early- and middle-stage mouse models, the corresponding CE-infected and normal control serum samples were collected, and silver nanoparticles (AgNPs) were utilized as the substrate to obtain SERS spectra. The early- and middle-stage discriminant models were developed using a support vector machine, with diagnostic accuracies of 91.7% and 95.7%, respectively. Furthermore, by analyzing the serum SERS spectra, some biomarkers that may be related to early CE were found, including purine metabolites and protein-related amide bands, which was consistent with other biochemical studies. Thus, our findings indicate that label-free serum SERS analysis is a potential early-stage CE detection method that is promising for clinical translation.

Evaluation of immunochromatography method in the diagnosis of cystic echinococcosis

Aims: Echinococcus granulosus is the causative agent of hydatid cyst, or cystic echinococcosis (CE), with its current name. Echinococcus granulosus is a zoonotic cestode; commonly found in humans and farm animals. In cystic echinococcosis infection, transmission occurs by oral ingestion of parasite eggs excreted in infected dog feces. The larval form is responsible for the formation of slowly growing cysts in the organs and tissues of mammals such as humans, sheep, goats and cattle. In this study, it was aimed to compare indirect hemagglutination (IHA) and immunochromatographic (ICT) methods from the sera of patients with suspected CE and to evaluate serological tests based on imaging and clinical diagnosis.&#x0D; Methods: Between 31 October 2022 and 31 January 2023, blood samples of 95 patients with suspected CE from different units of our hospital and for whom IHA was routinely requested were included in the study prospectively. VIRAPID® Hydatidosis (Vircell, Granada, Spain) test using the immunochromatographic method and ELI.H.A. Echinococcus (ELITech Microbio, France) test was studied in accordance with the manufacturer’s instructions.&#x0D; Results: Based on clinical and imaging methods of 95 patients included in our study, 64 (63.1%) were diagnosed with hydatid cyst. Based on imaging and clinical diagnosis; sensitivity, specificity, positive predictive value and negative predictive values were calculated as 81.3%, 96.8%, 98.1%, 71.4% for the IHA test, and were calculated as 75.0%, 93.5%, 96.6%, 64.4% for the ICT test, respectively. Good agreement was found between the two tests (percent agreement=68.0%; kappa value=0.682; p

The Importance of Indirect Hemagglutination Test in the Diagnosis of Cystic Echinococcosis: Serological– Histopathological Correlation

The Importance of Indirect Hemagglutination Test in the Diagnosis of Cystic Echinococcosis: Serological– Histopathological Correlation

Comparison of Native Hydatid Cyst Fluid (HCF), Lyophilized HCF, Antigen B (AgB) and Lyophilized AgB (LAgB) Originated from Echinococcus granulosus Sensu Stricto for Sero-Diagnosis of Active, Transitional and Inactive Human Liver Cystic Echinococcosis.

Cystic echinococcosis (CE) is an important zoonotic parasitic disease caused by the larval stage or metacestode of the tapeworm Echinococcus granulosus sensu lato. Due to treatment protocols for different liver cysts, diagnosis of cyst stages is very important. Different antigens have been used for CE diagnosis. However, each one is more sensitive and effective for the diagnosis of specific CE stages is not known well. We aimed to compare Native Hydatid Cyst Fluid (HCF), Lyophilized Hydatid Cyst Fluid (LHCF), antigen B (AgB) and Lyophilized antigen B (LAgB) originated from E. granulosus sensu stricto (G1-G3) genotype, for sero- diagnosis of active, transitional and inactive human liver CE using ELISA technique. The HCF was collected aseptically from liver CE cysts of sheep slaughtered from 2018 to 2019 in Shiraz slaughterhouse, Southern, Iran. The cysts were characterized by PCR and sequencing for genotype specification. Four types of antigens were used: HCF, LHCF, AgB and LAgB originated from E. granulosus sensu stricto (G1-G3) genotype. Thirty-three serum samples from active, transitional, and inactive human cysts were collected. Overall, 48 samples from other parasitic diseases and 60 samples from healthy subjects as negative controls were checked using four antigens by ELISA method. The best diagnostic sensitivity with 96.97% was observed by anti-LHCF IgG ELISA test. The best specificity with 95.37% was observed in ELISA test using LAgB. Simultaneous test of sera with anti-LHCF IgG ELISA and anti-LAgB IgG ELISA would be the best in the diagnosis of human liver cystic echinococcosis.

Ultrasound identification of hepatic echinococcosis using a deep convolutional neural network model in China: a retrospective, large-scale, multicentre, diagnostic accuracy study

Ultrasonography is the most widely used technique to diagnose echinococcosis; however, challenges in using this technique and the demand on medical resources, especially in low-income or remote areas, can delay diagnosis. We aimed to develop a deep convolutional neural network (DCNN) model based on ultrasonography to identify echinococcosis and its types, especially alveolar echinococcosis. This retrospective, large-scale, multicentre study used ultrasound images from patients assessed at 84 hospitals in China, obtained between Jan 1, 2002, and Dec 31, 2021. Patients with a diagnosis of cystic echinococcosis, alveolar echinococcosis, or seven other types of focal liver lesions were included. We tested ResNet-50, ResNext-50, and VGG-16 as the backbone network architecture for a classification DCNN model and input the perinodular information from the ultrasound images. We trained and validated the DCNN model to diagnose and classify echinococcosis using still greyscale ultrasound images of focal liver lesions in four stages: differentiating between echinococcosis and other focal liver lesions (stage one); differentiating cystic echinococcosis, alveolar echinococcosis, and other focal liver lesions (stage two); differentiating cystic echinococcosis, alveolar echinococcosis, benign other focal liver lesions, and malignant focal liver lesions (stage three); and differentiating between active and transitional cystic echinococcosis and inactive cystic echinococcosis (stage four). We then tested the algorithm on internal, external, and prospective test datasets. The performance of DCNN was also compared with that of 12 radiologists recruited between Jan 15, 2022, and Jan 28, 2022, from Qinghai, Xinjiang, Anhui, Henan, Xizang, and Beijing, China, with different levels of diagnostic experience for echinococcosis and other focal liver lesions in a subset of ultrasound data that were randomly chosen from the prospective test dataset. The study is registered at ClinicalTrials.gov (NCT03871140). The study took place between Jan 1, 2002, and Dec 31, 2021. In total, to train and test the DCNN model, we used 9631 liver ultrasound images from 6784 patients (2819 [41·7%] female patients and 3943 [58·3%] male patients) from 87 Chinese hospitals. The DCNN model was trained with 6328 images, internally validated with 984 images, and tested with 2319 images. The ResNet-50 network architecture outperformed VGG-16 and ResNext-50 and was generalisable, with areas under the receiver operating characteristic curve (AUCs) of 0·982 (95% CI 0·960-0·994), 0·984 (0·972-0·992), and 0·913 (0·886-0·935) in distinguishing echinococcosis from other focal liver lesions; 0·986 (0·966-0·996), 0·962 (0·946-0·975), and 0·900 (0·872-0·924) in distinguishing alveolar echinococcosis from cystic echinococcosis and other focal liver lesions; and 0·974 (0·818-1·000), 0·956 (0·875-0·991), and 0·944 (0·844-0·988) in distinguishing active and transitional cystic echinococcosis from inactive echinococcosis in the three test datasets. Specifically, in patients with the hepatitis B or hepatitis C virus, the model could distinguish alveolar echinococcosis from hepatocellular carcinoma with an AUC of 0·892 (0·812-0·946). In identifying echinococcosis, the model showed significantly better performance compared with senior radiologists from a high-endemicity area (AUC 0·942 [0·904-0·967] vs 0·844 [0·820-0·866]; p=0·027) and improved the diagnostic ability of junior, attending, and senior radiologists before and after assistance with AI with comparison of AUCs of 0·743 (0·714-0·770) versus 0·850 (0·826-0·871); p<0·0001, 0·808 (0·782-0·832) versus 0·886 (0·864-0·905); p<0·0001, and 0·844 (0·820-0·866) versus 0·870 (0·847-0·890); p=0·092, respectively. The DCNN model was shown to be accurate and robust, and could improve the ultrasound diagnostic ability of radiologists for echinococcosis and its types for highly endemic and remote regions. National Natural Science Foundation of China and National Key Research & Development Program of China. For the Chinese translation of the abstract see Supplementary Materials section.

Parasite-Derived MicroRNAs: Potential Alternative Targets for Laboratory Diagnosis of Cystic Echinococcosis

Cystic Echinococcosis (CE) is a type of zoonotic infection that can be caused by a specific form of a parasite called Echinococcus granulosus sensu lato. Mostly, imaging techniques are utilized to diagnose CE. Serological tests are used only when imaging findings are atypical. Additionally, laboratory assays including direct microscopy and PCR are used in the confirmation of diagnosis after treatment though obtained negative results with these tests cannot be ruled out the diagnosis. Specific miRNAs produced by the parasite could be used as markers to diagnose and monitor CE. This research focuses on investigating the diagnostic potential of parasite-derived miRNAs compared to the presence of protoscolex in animal-derived hydatid cyst samples. Accordingly, egr-let-7-5p, egr-miR71-5p and egr-miR-9-5p were found to be positive in 26, 25 and 11 out of 30 samples (86.6%, 83.3% and (36.6%), respectively. There was no relationship between protoscolex presence and detection of either egr-let-7-5p or egr-miR-9-5p (p&gt;0.05). On the other hand, egr-miR71-5p positivity was found statistically significant compared with protoscolex presence (p=0.04). As a result, egr-miR-71 are a promising potential target for the diagnosis of CE. Additional research is necessary to assess the diagnostic value of miRNAs in CE using a larger group of samples.

Application and evaluation of native antigen B from Echinococcus granulosus sensu stricto and E. canadensis alone or mixture for serodiagnosis of human G1-G3 and G6/G7 genotypes cystic echinococcosis sera, using ELISA and Western blotting.

Cystic echinococcosis (CE) is one of the most important helminthic diseases in the world with different genotypes distribution. The application of specific genotype antigens together with sera from patients with specific cyst genotypes have not been reported, so far. The present study aimed to apply and evaluate native AgB from Echinococcus granulosus sensu stricto (Eg) and Echinococcus canadensis (Ec) alone or mixture for serodiagnosis of human G1-G3 and G6/G7 genotypes cystic echinococcosis sera, using ELISA and Western blotting. A total of 47 human sera along with 47 human CE cysts were collected. CE genotypes were determined. Native AgB were prepared from E. granulosus s.s and E. canadensis genotypes. ELISA and Western blot were performed on human specific G1-G3 and G6/G7 genotypes sera. Species specific native AgB were used alone or mixed. The sensitivity of ELISA using alone and mixed 1Eg-1Ec, 1Eg-2Ec, and 2Eg-1Ec of native AgB from E. granulosus s.s and E. canadensis genotypes for human G1-G3 sera were 92.10, 89.47, 97.37, 100, and 100%, respectively; while using AgBs, alone and mixed for human G6/G7 sera were 100%. The sensitivity of Western blotting using native AgB of E. granulosus s.s and E. canadensis genotypes alone and mixed 2Eg-1Ec were 78.95% and 100% for human G1-G3 and G6/G7 genotypes sera, respectively. The mixture of AgB from Echinoccus granulosus sensu stricto and Echinococcus canadensis genotypes increased ELISA sensitivity for the diagnosis of human CE. Preparation and application of native AgB from specific and prevalent genotypes of CE in endemic regions is recommended.

Ultrasound-Based Prevalence of Cystic Echinococcosis in the Samarkand Region of Uzbekistan: Results from a Field Survey.

Cystic echinococcosis (CE) is a zoonosis caused by Echinococcus granulosus. Uzbekistan is endemic for CE, but estimates of disease burden are lacking. We present findings from a cross-sectional, ultrasound-based survey evaluating the prevalence of human CE in the Samarkand region, Uzbekistan. The survey was conducted between September and October 2019 in the Payariq district, Samarkand. Study villages were selected based on sheep breeding and reported human CE. Residents aged 5-90 years were invited to receive a free abdominal ultrasound examination. The WHO Informal Working Group on Echinococcosis classification was used for cyst staging. Information regarding CE diagnosis and treatment was collected. Of 2,057 screened subjects, 498 (24.2%) were male. Twelve (0.58%) had detectable abdominal CE cysts. In total, five active/transitional (N = 1 CE1, N = 1 CE2, N = 3 CE3b) and 10 inactive cysts (N = 8 CE4, N = 2 CE5) were identified. Two participants had cystic lesions with no pathognomonic features of CE and were given a 1-month course of albendazole for diagnostic purposes. Twenty-three additional individuals reported previous surgery for CE in the liver (65.2%), lungs (21.6%), spleen (4.4%), liver and lungs (4.4%), and brain (4.4%). Our findings confirm the presence of CE in the Samarkand region, Uzbekistan. Additional studies are needed to assess the burden of human CE in the country. All patients with a history of CE reported surgery, even though most cysts found during the current study were inactive. Therefore, it appears there is a lack of awareness by the local medical community of the currently accepted stage-specific management of CE.

IgG glycomic profiling identifies potential biomarkers for diagnosis of echinococcosis

Echinococcosis caused by larval stage of the genus Echinococcus, is a serious and potentially fatal parasitic zoonosis distributed globally. The two types of the disease in human are cystic echinococcosis (CE) and alveolar echinococcosis (AE). As the biological and encysting characteristics of the parasite, early diagnosis remains to address. In the present study, we demonstrate the value of Immunoglobulin G (IgG) glycome as a potential diagnostic biomarker for echinococcosis. Serum IgG glycome profiles were analyzed by ultra-performance liquid chromatography in a cohort comprised of 127 echinococcosis patients, of them 98 were diagnosed as CE and 29 as AE. IgG N-glycome analysis in pretreatment serum of echinococcosis patients presents 25 glycans and 64 derived traits. Compared with IgG glycans of healthy control group, neutral glycans, fucosylation and agalactosylated N-glycans increased while sialylation and galactosylation decreased in echinococcosis patients. Combined with a machine-learning-based approach, we built three biomarker combinations to distinguish CE, AE and healthy controls. Meanwhile, galactosylation, sialylation and A2BG2S1 in IgG glycan profiles were evidently associated with different types of CE (from CE1 to CE5). Our findings suggest that the alterations in IgG N-glycome may be of value in CE and AE diagnosis and follow-up CE disease progress. The role of IgG N-glycans as diagnostic biomarker remains to be verified in future study.

Molecular characterization of cattle and sheep isolates of Echinococcus granulosus from Elazig province in Turkey and expression analysis of the non-coding RNAs, egr-miR-7, egr-miR-71 and egr-miR-96

Cystic Echinococcosis (CE) is a common zoonotic disease seen in human and animals worldwide, caused by the larval form of Echinococcus granulosus. In this study, E. granulosus s.l. species and haplotypes were determined in hydatid cysts isolated from cattle and sheep, and the expression levels of egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were compared in different cyst structures. A total of 82 (cattle, n = 41; sheep, n = 41) hydatid cyst isolates (germinal membranes and/or protoscoleces) were collected from a slaughterhouse in Elazig province of Turkey. After mt-CO1 gene sequences were made, 81 out of 82 hydatid cyst isolates were determined as E. granulosus s.s. (G1 and G3), while an isolate of cattle origin was determined as Echinococcus canadensis (G6/7). A total of 26 nucleotide polymorphisms and 29 haplotype groups were identified in the samples. miRNA expressions in germinal membranes of sterile cysts and germinal membrane and protoscoleces of fertile cysts were investigated by qRT-PCR and Real Time PCR analyses. It was determined that miRNAs were expressed at high levels in 79.31% of the 29 haplotype groups and at low levels in the remaining 10.34%. In 10 fertile samples of sheep origin, egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were found to be 44, 168, and 351-fold higher in expression, respectively, in the germinal membrane compared to the protoscoleces. Especially egr-miR-96 may have the potential to be used as biomarkers in the diagnosis of active CE.

Molecular characterization and immunological properties of Echinococcus granulosus sensu stricto (G1) ADK1 and ADK8.

Adenylate kinases (ADKs) are one of the important enzymes regulating adenosine triphosphate (ATP) metabolism in Echinococcus granulosus sensu lato. The objective of the present study was to explore the molecular characteristics and immunological properties of E. granulosus sensu stricto (G1) adenylate kinase 1 (EgADK1) and adenylate kinase 8 (EgADK8). EgADK1 and EgADK8 were cloned and expressed, and the molecular characteristics of EgADK1 and EgADK8 were analyzed through different bioinformatics tools. Western blotting was used to examine the reactogenicity of recombinant adenylate kinase 1 (rEgADK1) and recombinant adenylate kinase 8 (rEgADK8) and to evaluate their diagnostic value. The expression profiles of EgADK1 and EgADK8 in 18-day-old strobilated worms and protoscoleces were analyzed by quantitative real-time PCR, and their distribution in 18-day-old strobilated worms, the germinal layer, and protoscoleces was determined by immunofluorescence localization. EgADK1 and EgADK8 were successfully cloned and expressed. Bioinformatics analysis predicted that EgADK1 and EgADK8 have multiple phosphorylation sites and B-cell epitopes. Compared with EgADK8, EgADK1 and other parasite ADKs have higher sequence similarity. In addition, both cystic echinococcosis (CE)-positive sheep sera and Cysticercustenuicollis-infected goat sera could recognize rEgADK1 and rEgADK8. EgADK1 and EgADK8 were localized in protoscoleces, the germinal layer, and 18-day-old strobilated worms. EgADK1 and EgADK8 showed no significant difference in their transcription level in 18-day-old strobilated worms and protoscoleces, suggesting that EgADK1 and EgADK8 may play an important role in the growth and development of E. granulosus sensu lato. Since EgADK1 and EgADK8 can be recognized by other parasite-positive sera, they are not suitable as candidate antigens for the diagnosis of CE.

Multimodal Treatment of Cystic Echinococcosis

Cystic echinococcosis is a zoonotic disease that is caused by the larval stages of cestodes species and belongs to the genus Echinococcus. Echinococcus granulosus (E. granulosus) causes cystic echinococcosis which is global and wider in its distribution than alveolar echinococcosis. Endemic areas of E. granulosus are Russia, Eastern Europe, the Middle East, China, and South America based on data from the World Health Organization (WHO). Incidence rates are 50 per 100,000 person-years. The life cycle of E. granulosus requires both an intermediate host and a definitive host. A human accidentally becomes an intermediate host. Upon infection, cyst formation mostly occurs in the liver (70%). At the first, the infection is usually asymptomatic. Diagnosis of cystic echinococcosis can be done by imaging techniques (ultrasound or CT/MRI), serum serologic testing for antibodies against hydatid antigens, and immunologic testing. In general, there are four different management modalities for cystic echinococcosis, such as surgery, percutaneous therapy surgery, chemotherapy, and watchful waiting.

Seropositivity Pattern of Human Cystic Echinococcosis at a Tertiary Care Hospital of India.

Cystic echinococcosis (CE), even after several control measures, causes significant morbidity throughout the world. Besides imaging investigation technology, the serological tests are essential for both diagnosis and management of this slowly progressive disease. The present study was a hospital-based retrospective study that examined the seropositivity rate for Echinococcus granulosus sensu lato antibody in patients suspected of CE at our tertiary health care center over 8 years from 2013 to 2020. Records of new visits to hospital/clinics and associated hospital discharge constituted the denominator of calculation. All samples were tested using commercially available indirect immunoglobulin G enzyme-linked immunosorbent assay kit. A total of 925 suspected patients with a clinical diagnosis of CE were screened. The age group that commonly tested positive for CE was 20 to 39 years, and liver was the predominant organ found to be affected. The seropositive rate was 41.2%. On further year-wise analysis, it was observed that the seropositivity rate had significantly declined from 61.4% in 2013 to 33.8% in 2020. This study clearly showed that there is a by 27.6% decline of CE seropositivity rate in 8 years. This declining rate may be attributed to improved socioeconomic status and better implementation of health programs.

In silico study to predict promiscuous peptides for immunodiagnosis of cystic echinococcosis.

Cystic echinococcosis (CE), caused by Echinococcus granulosus, is a major zoonotic disease that causes significant human morbidity and mortality. This cosmopolitan disease is difficult to diagnose, treat, and control. So far, crude extracts of hydatid cyst fluid containing antigen B or antigen 5 have been used as the primary antigenic source for its immunodiagnosis. The main issue is that it reacts with sera from people infected with other helminths. There is currently no standard, specific, or sensitive test for disease diagnosis, and no human vaccine has been reported. Considering the need for efficient immunization and/or immunodiagnosis, six E. granulosus antigens, antigen 5, antigen B, heat shock proteins such as Hsp-8 and Hsp-90, phosphoenolpyruvate carboxykinase, and tetraspanin-1, were chosen. Using various in silico tools, T cell and B cell epitopes (promiscuous peptides) were predicted by targeting antigen 5, antigen B, heat shock proteins such as Hsp-8 and Hsp-90, phosphoenolpyruvate carboxykinase, and tetraspanin-1. There are twelve promiscuous peptides with overlapping human leukocyte antigen (HLA) class-I, class-II, and conformational B cell epitopes. Such immunodominant peptides could be useful as subunit vaccines. Furthermore, six peptides specific for E. granulosus were also discovered, which may prove to be important markers in the diagnosis of CE, potentially preventing misdiagnosis and mismanagement. These epitopes may be the most important vaccine targets in E. granulosus because they have the most promiscuous peptides and B cell epitopes, as well as the highest affinity for different alleles, as determined by docking scores. However, additional research using in vitro and in vivo models is undertaken.

Diagnostic Efficiency of Heat Shocked Protoscoleces Extract Antigens for Human Cystic Echinococcosis by ELISA

      Diagnosis of cystic echinococcosis is complex and has to be confirmed by the combination of immunological tests and imaging techniques. In this study heat shock proteins were induced and their immunoreactivity was assessed by ELISA. Sera were collected from 34 hydatid patients who were admitted to the Rizgary Teaching Hospital through October 2013 to July 2017, in addition to 29 healthy donors and 18 non-hydatid cases. For heat shock response, two batches of 25000 protoscoleces (Ps) were incubated separately at 42°C and 45 °C for 4 hours. Heat treated and normal Ps were disrupted and the extracts were divided into two parts. One part was directly used as source of antigens (PE, PE42 and PE45) and the other one was partially purified on Sephadex G150. The immunoreactivity of these antigens, as well as hydatid fluidwas assessed by ELISA. The cutoff value to differentiate positive from negative sera was established by receiver operating characteristic (ROC) analysis. Two peaks of PE42 and three peaks of PE45 resulted by Sephadex G150. Extracts of 42ºC treated Ps resulted in two protein peaks and were used as PE42P1 and PE42P2 antigens. For 45°C treated Ps, the chromatography patterns resulted in three protein peaks and were used as PE45P1, PE45P2 and PE45P3 antigens. Highest rates of sensitivity, specificity and diagnostic accuracy were detected with PE42P2 (91.2%) and PE45P2 (91.2%). Sensitivity of ELISA was consistent for liver cysts with all applied antigens. Hydatid antigens extracted from heat treated Ps markedly raised the sensitivity of ELISA to detect anti-hydatid IgG.

Can Echinococcus granulosus-Derived MicroRNAs be Biomarkers for Diagnosis and Follow-up of Cystic Echinococcosis Patients?

Cystic echinococcosis (CE) is a neglected tropical disease caused by the larval stages of Echinococcus granulosus (E. granulosus). MicroRNAs (miRNAs) are small noncoding RNAs acting as mediators in host-parasite interaction. Recently, numerous studies have been conducted on miRNAs in infectious diseases; however, little data are available about the role of miRNAs in pathogenesis and early diagnosis of CE. The current study evaluated the expression of four E. granulosus-derived miRNAs, including egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71 in fibrotic and healthy liver tissues of 31 CE patients with active and inactive hydatid cysts by qRT-PCR. Of the 31 patients, 48.4% had active cysts (CE1 and CE2), while the remainder had transitional (16.1%) and inactive (35.5%) CE types cysts. The qRT-PCR analysis revealed a significant increase of 11.2, 9.91, 6.2, and 13.1-fold in the fibrotic tissue group for egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71, respectively. Among these miRNAs, egr-miR-125-5p exhibited the highest area under the curve (AUC) value of 0.8050 for predicting liver fibrosis. Our findings provide new data about the role of E. granulosus-derived miRNAs in pathogenesis of CE. The high AUC of egr-miR125,5p reflecting the possibility of using egr-miR125,5p as biomarker in CE diagnosis. Further studies on serum of CE patients are needed to confirm the potential role of circulating egr-miR-2a-3p and egr-miR-125-5p in the early diagnosis of CE.