Molecular Characterization of Echinococcus granulosus isolates from Human Cases Using Gold Nanoparticles-Based DNA Microarray with Silver Enhancement Simple Colorimetric Technology (original) (raw)
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Genetic Fingerprint of Unilocular Hydatidosis in Egyptian Camels and Humans Using Nested PCR
The pulmonary and hepatic hydatid cyst fluids were collected from 540 slaughtered camels and 5 human cases in Qalyubia Governorate, Egypt. The prevalence of infection of cystic echinococcosis among camels was 120 (22.2%). The fertility rates of the isolated cysts form camels and humans were 64.5 and 100%, respectively. A nested polymerase chain reaction was used for amplification of mitochondrial NADH 1 gene of Echinococcus granulosus complex in fertile cysts obtained from camels and humans, respectively. Two pairs of primers (EGL1 and EGR2) and (EGL3 and EGR4) were used in 2 amplification steps. First, the outer pair of primer originated from a highly conserve region of NADH1 gene generate a primary 435 bp PCR product. Second, a pair of internal (nested) primer (EGL3 and EGR4), designed to the annealing site of primers (EGL1 and EGR2) yield similar diagnostic amplified DNA bands of molecular size marker at 276 bp in all examined cysts obtained from camels and humans indicating a zoonotic relationship. This study confirms similar fingerprinting patterns of Echinococcus granulosus complex in camels and humans in Qalyubia Governorate, Egypt. Nested PCR for diagnosis of E. granulosis had been used for the first time in Egypt, as far as we know.
Evaluation of crude Antigens for Serological Diagnosis of Hydatidosis in Man and Camel in Sudan
2016
Evaluation of crude Antigens for Serological Diagnosis of Hydatidosis in Man and Camel in Sudan. Adam Alfaki Mohammad Albadawi*, Mohammad Eltayeb Ahmed, Nawal Tagelsir M. Osman Mohammad Elowani, Teaser Elameen,Asim Abdelrahman Daffalla, Mohammad Baha Eldin Saad. Parasitology Department, National Public Health Laboratory, Ministry of Health, Republic of Sudan. Parasitology Department, National Health Laboratory, Ministry of Health, Republic of Sudan. Molecular Biology Department, National Health Laboratory, Republic of Sudan. Parasitology Department, Faculty of medical laboratory Science & Technology, Sudan University of Science & Technology, Republic of Sudan. Parasitology Department, Faculty of Medical Veterinary, Sudan University of Science & Technology, Republic of Sudan. Research Department, Faculty of Science, Abaha University Kingdom Saudi Arabia. Parasitology Department, Faculty of medical laboratory Science, Omdurman Ahlia University, Republic of Sudan.
2020
Examination of 528 (450 males and 78 females) dromedary camels slaughtered at Cairo abattoir revealed that a total of (93) 17.6 % were infected with hydatidosis. Post mortem examination revealed that infection was restricted only in the lungs and the liver of infected camels. Among the 93 hydatidosis infected camels, lungs were the most frequently infected 88 (94.623%) compared with liver 5 (5.376%). ELISA test using partially crude antigen and purified antigen is important for the early diagnosis of cystic echinococcosis as most cases in the early stages of infection are asymptomatic. Sensitivity of ELISA using the crude antigen was 82.758% while sensitivity of the partially purified antigen was 79.310 %. On the other hand specificity of the crude antigen was estimated as 62.5 % and specificity of partially purified antigen as 75.0 %.
Journal of Parasitic Diseases, 2014
Hydatid Disease is the name given to the condition caused by the zoonotic tapeworm Echinococcus granulosus. The tapeworm spends most of its adult life in the intestine of its definitive host, namely canids and in particular the dog. The tapeworm eggs become voided in the canids' faeces and as a result of ingesting the eggs, infection passes to the intermediate host, commonly herbivores while grazing. However, humans can become accidentally infected and hydatid cysts may develop throughout the body. During April 2010-February 2014, a total 198 camels, which had been sent to the abattoir, the daily number of hydatid infected livers and lungs of camels slaughtered at Tabriz abattoir were recorded. To be sure about the validity of recorded data, observed data were collected daily. Approximately 29 (14.64 %) of camels were infected according to this survey. Age wise, the prevalence of infection in young animals (under the age of 5 years) was 4 (2.02 %), whereas in animals between 5 and 10 years and over, the prevalence of infection was 11 (5.55 %) and 14 (7.07 %) respectively. Sex wise, female animals had a higher prevalence with 17 (19.76 %) cases in camels, whereas in the males, there were 12 (10.71 %) cases in camels. There was a notable difference found in our study between male and female animals (P \ 0.05). Infections were recorded in two visceral organs with the Lungs having the highest prevalence in camels, and the liver had low infected in camels. The results of this study suggest that infection of camels with hydatid cyst is common in Tabriz, Iran and that this may constitute economic and health problems in the meat industry.
The objectives of the present study were to investigate strain identification of Echinococcus granulosus infecting camel and human in Qalyubia, Egypt. Therefore partial sequences were generated after gel purification of nested PCR amplified products of mitochondrial NADH 1gene of Echinococcus granulosus complex. Sequences were further examined by sequence analysis and subsequent phylogeny to compare these sequences to those from known strains of E.granulosus circulating globally and retrieved from GenBank. All isolates are homologous to the camel strain, E. canadensis (G6) genotype. Nucleotide mutations generate polymorphism at position of 275 nucleotide, where a thymine replaced a cytosine and at the levels of 385 and 386 nucleotides, where two cytosine substituted a guanine and a thymine respectively. KF815488 Egypt showed typical identity (99.5%) with JN637176 Sudan, HM853659 Iran, AF386533 France and AJ237637 Poland with 0.5% diversion.. Phylogenetic analysis showed a robust tree clustering all isolates with sequences belonging to the camel genotype (G6) variant with strong bootstrap values at relevant nodes and the evolutionary distance between groups is very short. There are two mutations in the sequences of amino acids at the position of 92, where an Alanine is changed to a Valine and at the position of 129, where a Valine is transformed to a Proline. Our record of a single genotype determined a strain which could be incriminated for camel and human infectivity and responsible for its persistence in the endemic areas. Such epidemiological data could guide the application of efficient control strategies of hydatidosis in Egypt
CHARACTERISTICS OF HYDATID CYST OF CAMEL STRAIN OF ECHINOCOCCUS GRANULOSUS IN ASSIUT
minia.edu.eg
100 camels were examined for hydatid cyst and revealed that the prevalence rate of cystic hydatidosis among camels in Assiut Governorate was 9% .The lung was the most suitable place for cystic infection in camel than the liver. The fertility rate of the examined cysts was 80%. Hydatid cyst wall was better stained with PAS. Best carmine and Giemsa stains were very rapid effective methods for visualization of hydatid elements. Scanning electron microscope was used for the detection of ultrastructure features of hydatid sand. It revealed that camel strain has some specific characters including large sized rostellar hooks and presence of separation line between the scolex and the neck in the evaginated protoscolices. In top view the rostellum appeared to be formed of 3 successive concentric rings becoming smaller as they go upwards. This was shown for the first time in the present study. SDS PAGE was used for characterization of hydatid cyst fluid, protoscolices and laminated layer antigens separately. It revealed that protoscolices and laminated layer are the source of proteins in the hydatid fluid. Many protein bands ranged from 150-6 kD were detected by different stains. The pattern of these bands indicated that camel's hydatid cyst components differ from that of sheep and cattle.
MOLECULAR STUDY OF HYDATID CYSTS IN HUMAN, SHEEP AND DONKEYS AT BASRA CITY, SOUTHERN IRAQ
Biochem. Cell. Arch., 2019
Hydatid disease or Echinoccosis is one of the serious public health problems. This study designed for molecular investigation of Hydatid cysts in different hosts animals (Sheep, donkey) and human in Basra city, as well as molecular characterization by PCR technique to determine the specific gene for human (ND1, COI, CO1), sheep (G6-7, sh4-1, COI) and donkey (COI, CO1). Then, gene sequencing to recognize the intraspecific variation of Echincoccus in relation to the host. In the current study, 818 sheep were examined and the number of sheep infected with hydatid cysts was 271. Donkey samples (10) were brought to the animal field of Veterinary Medicine at the University of Basra and the number of infected with hydatid cysts was two in the period from September 2017 to March 2018. The hydatid cysts were collected from the human after surgery at Al-Sadr Teaching Hospital where 21. The results were higher in female (15) than male (6) in the same period.
Veterinary World, 2020
Background: Cystic echinococcosis (CE), a zoonotic disease that affects animal and human health, is of increasing economic importance due to high morbidity rates and high economic losses in the livestock industry. Aim: The present study was conducted to purify the antigen from hydatid cyst fluid (HCF) with high diagnostic efficacy of camel hydatidosis using indirect enzyme-linked immunosorbent assay (ELISA). Materials and Methods: The HCF antigen was purified using Sephacryl S-300 column chromatography. Characterization of fractions was performed using reducing and non-reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Further, antibodies against Echinococcus granulosus cysts in camel serum were detected using indirect ELISA. Results: The purification process resulted in three fractions of antigens: FI, FII, and FIII. Indirect ELISA showed that higher diagnostic efficacy was observed in FI than in FII and FIII. Indirect ELISA, in which FI was utilized, showed 88% sensitivity and 91.7% specificity. Non-reducing SDS-PAGE showed that FI had two bands of molecular weights 120 and 60 kDa. Western blot analysis of FI demonstrated that 60, 38, and 22 kDa were antigenic bands when reacted with naturally infected camel sera with E. granulosus cysts. Using indirect ELISA, F1 recorded an infection percentage of 81.7% in randomly collected camel serum samples. Conclusion: FI is a promising antigen for accurate diagnosis of camel CE using indirect ELISA.
Veterinary World, 2021
Background and Aim: Cystic echinococcosis (CE)/hydatidosis is one of the most prevalent neglected zoonotic diseases. It is initially asymptomatic and does not produce any clinical signs until the cyst becomes enlarged, causing localized pressure on internal organs and tissues. Therefore, the detection of Echinococcus granulosus antibodies is highly essential. This study evaluated the antigens of hydatid cyst fluid, protoscoleces, and germinal layers for efficient immunological diagnosis of CE in humans and camels. Materials and Methods: Hydatid cyst fluid (FLc), protoscoleces (Psc), and the germinal layer (GLc) antigens were prepared from camel-lung hydatid cysts. In the same way, hydatid cyst fluid (FLh) and protoscoleces (Psh) antigens from human-liver cyst aspirate were produced. The comparative immunodiagnostic efficacy of the prepared antigens was verified using indirect enzyme-linked immunosorbent assay (ELISA), SDS-PAGE, and immunoblotting. Results: ELISA proves that FLc and GLc antigens were higher than FLh and Psh antigens. This shows that binding reactivity in naturally infected human sera, camel sera, and Psc is the most potent, exhibiting 100% sensitivity with 78.26% and 76.47% specificity in camel and human sera, respectively. The CE prevalence using diagnostic Psc was 54.79% and 61.32% in tested human and camel sera, respectively. The electrophoretic profiles of all shared antigens showed similarities at 52, 41, and 22 kDa. Immunoblotting demonstrated common immune-reactive bands in all antigen types at 52 and 41 kDa against positive human and camel sera. Conclusion: This immunological study introduces camel hydatid cyst Psc as a potent diagnostic antigen and new immune-reactive fractions of 52 and 41 kDa for diagnosing hydatidosis in humans and camels.
2013
4 Abstract: Hydatidosis is highly public health problem and economic importance worldwide. A cross-sectional survey of camel hydatidosis was carried out on 421 camels from October 2012 to May 2013 in Ayssaita district. The main objectives of this study were to estimate the prevalence and identify the potential risk factors to acquire hydatidosis infection in camels slaughtered in backyard in pastoral area. The overall prevalence of camel hydatidosis was found to be 34.20% (95%CI: 29.65, 38.75). Based on the potential risk factors, the likelihood of acquiring camel hydatidosis was higher in female than male (OR = 1.73, 95% CI: 1.11, 2.69, P = 0.02), in old than young (OR = 5.75, 95% CI: 2.88, 11.49, P < 0.0001), in lean than good body condition (OR = 4.5, 95% CI: 2.15, 9.13, P <0.0001). Sources of water and feeding habit of camels was also found to be statistically significant (P<0.05). Of the 144 camels positive, 47.90% had cysts in the lungs only, 20.80% in the liver only,...