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Fulltext IgG4 detection of Echinococcus granulosus paramyosin is a useful diagnostic test for human hydatidosis

Moghadam ZK, Ghaffarifar F, Khalilpour A, Abdul Aziz F, Saadatnia G, Noordin R

Clin Vaccine Immunol, 2013 Apr;20(4):501-5.
PMID: 23365208 DOI: 10.1128/CVI.00019-13

Hydatidosis is a public health problem in many parts of the world, and improvement in diagnosis of the disease is still being pursued. Protoscoleces of Echinococcus granulosus were isolated from hydatid cysts collected from naturally infected sheep slaughtered in abattoirs in Iran. Sonicated extract of protoscolex was subjected to two-dimensional gel electrophoresis and Western blot analysis. Primary antibodies were from serum samples from 130 hydatidosis patients, 38 individuals infected with other parasitic infections, and 30 healthy people, whereas peroxidase (HRP)-conjugated anti-human IgG and IgG4 were used as secondary antibodies. The recombinant form of the identified protein was produced and tested for its sensitivity and specificity for the detection of human hydatidosis. An antigenic band of ∼60 kDa was found to be sensitive (82%) and specific (100%) for the detection of hydatidosis when probed with anti-human IgG4-HRP, while the sensitivity and specificity were 33 and 100%, respectively, with anti-human IgG-HRP. By mass spectrometry, the band was identified as protoscolex tegument paramyosin. The sensitivity and specificity of full-length paramyosin-recombinant protein in IgG4 blots were found to be 86 and 98%, respectively. In conclusion, IgG4 detection of Echinococcus granulosus paramyosin was found to be useful for the diagnosis of human hydatidosis.

Matched MeSH terms: Antibodies, Helminth/blood*
A filtration-based rapid test using a partially purified third-stage larval antigen to detect specific antibodies for the diagnosis of gnathostomiasis

Ma A, Wang Y, Liu XL, Zhang HM, Eamsobhana P, Yong HS, et al.

J Helminthol, 2019 Jan;93(1):26-32.
PMID: 29144215 DOI: 10.1017/S0022149X17001080

Human gnathostomiasis is an emerging food-borne parasitic disease caused by nematodes of the genus Gnathostoma. Currently, serological tests are commonly applied to support clinical diagnosis. In the present study, a simple and rapid filtration-based test, dot immune-gold filtration assay (DIGFA) was developed using a partially purified antigen of Gnathostoma third-stage larvae (L3). A total of 180 serum samples were tested to evaluate the diagnostic potential of DIGFA for gnathostomiasis. The diagnostic sensitivity and specificity were 96.7% (29/30) and 100% (25/25), respectively. The cross-reactivity with sera from other helminthiasis patients ranged from 0 to 4%, with an average of 1.6% (2/125). DIGFA using a partially purified L3 antigen was not only simple and rapid, but also more accurate than standard assays for the diagnosis of human gnathostomiasis. DIGFA may represent a promising tool for application in laboratories or in the field, without requiring any instrumentation.

Matched MeSH terms: Antibodies, Helminth/blood*
Fulltext Field evaluation of a rapid diagnostic test to detect antibodies in human toxocariasis

Lim PK, Yamasaki H, Mak JW, Wong SF, Chong CW, Yap IK, et al.

Acta Trop, 2015 Aug;148:32-7.
PMID: 25910623 DOI: 10.1016/j.actatropica.2015.04.011

Human toxocariasis which is caused mainly by the larvae of Toxocara canis and Toxocara cati, is a worldwide zoonotic disease that can be a potentially serious human infection. The enzyme-linked immunosorbent assay (ELISA) using T. canis excretory-secretory (TES) antigens harvested from T. canis larvae is currently the serological test for confirming toxocariasis. An alternative to producing large amounts of Toxocara TES and improved diagnosis for toxocariasis is through the development of highly specific recombinant antigens such as the T. canis second stage larva excretory-secretory 30 kDa protein (recTES-30). The aim of this study was to evaluate the sensitivity and specificity of a rapid diagnostic kit (RDT, named as iToxocara kit) in comparison to recTES-30 ELISA in Serendah Orang Asli village in Selangor, Malaysia. A total of 133 subjects were included in the study. The overall prevalence rates by ELISA and RDT were 29.3% and 33.1%, respectively, with more positive cases detected in males than females. However, no association was found between toxocariasis and gender or age. The percentage sensitivity, specificity, positive predictive value and negative predictive value of RDT were 85.7%, 90.1%, 80% and 93.2%, respectively. The prevalence for toxocariasis in this population using both ELISA and RDT was 27.1% (36/133) and the K-concordance test suggested good agreement of the two tests with a Cohen's kappa of 0.722, P<0.01. In addition, the followed-up Spearman rank correlation showed a moderately high correlation at R=0.704 and P<0.01. In conclusion, the RDT kit was faster and easier to use than an ELISA and is useful for the laboratory diagnosis of hospitalized cases of toxocariasis.

Matched MeSH terms: Antibodies, Helminth/blood
Brugia malayi infection in Meriones unguiculatus: antibody response to recombinant BmR1

Lim BH, Noordin R, Nor ZM, Rahman RA, Abdullah KA, Sinnadurai S

Exp Parasitol, 2004 Sep-Oct;108(1-2):1-6.
PMID: 15491542

BmR1 recombinant antigen has previously been shown to demonstrate high sensitivity and specificity in the serological diagnosis of brugian filariasis in humans. In this study, the pattern of recognition of antibody to BmR1 during Brugia malayi infection was investigated by employing Meriones unguiculatus as the experimental model. Thirty two gerbils were infected subcutaneously with 120 L(3); and two control groups each comprising 25 animals were employed. ELISA using BmR1 was used to detect filaria-specific IgG antibodies elicited by the gerbils; using sera collected from the day 1 until day 150 post-inoculation (p.i.). The results showed that BmR1 detected B. malayi infection in gerbils harboring adult worms irrespective of the presence of circulating microfilaria, and was exemplified by positive ELISA results in nine a microfilaraemic animals that harbored live adult worms. The initial time of the antibody recognition was at day 8 p.i. and the antibody titre showed some correlation with adult worm burden.

Matched MeSH terms: Antibodies, Helminth/blood
Fulltext High seroprevalence of echinococossis, schistosomiasis and toxoplasmosis among the populations in Babati and Monduli districts, Tanzania

Khan MB, Sonaimuthu P, Lau YL, Al-Mekhlafi HM, Mahmud R, Kavana N, et al.

Parasit Vectors, 2014;7:505.
PMID: 25388913 DOI: 10.1186/s13071-014-0505-7

The neglected tropical diseases, echinococcosis, schistosomiasis and toxoplasmosis are all globally widespread zoonotic diseases with potentially harmful consequences. There is very limited data available on the prevalence of these infections, except for schistosmiasis, in underdeveloped countries. This study aimed to determine the seroprevalence of Echinococcus multilocularis, Schistosoma mansoni, and Toxoplasma gondii antibodies in populations from the Monduli and Babati districts in Tanzania.

Matched MeSH terms: Antibodies, Helminth/blood*
Fulltext Lateral flow test using Echinococcus granulosus native antigen B and comparison of IgG and IgG4 dipsticks for detection of human cystic echinococcosis

Khalilpour A, Sadjjadi SM, Moghadam ZK, Yunus MH, Zakaria ND, Osman S, et al.

Am J Trop Med Hyg, 2014 Nov;91(5):994-9.
PMID: 25200268 DOI: 10.4269/ajtmh.14-0170

Cystic echinococcosis (CE) caused by infection with Echinococcus granulosus is of major concern for humans in many parts of the world. Antigen B was prepared from E. granulosus hydatid fluid, and Western blots confirmed eight batches showing a band corresponding to the 8-/12-kDa subunit with positive serum and no low-molecular mass band (< 15 kDa) with negative serum. The batches were pooled and used to prepare lateral flow immunoglobulin G4 (IgG4) and IgG dipsticks. Diagnostic sensitivity was determined using serum samples from 21 hydatidosis patients, and diagnostic specificity was established using sera from 17 individuals infected with other parasites and 15 healthy people. IgG4 dipstick had a diagnostic sensitivity of 95% (20 of 21) and a specificity of 100% (32 of 32). The IgG dipstick had a sensitivity of 100% (21 of 21) and a specificity of 87.5% (28 of 32). Thus, both IgG and IgG4 dipsticks had high sensitivities, but IgG4 had greater specificity for the diagnosis of human CE.

Matched MeSH terms: Antibodies, Helminth/blood
Fulltext Seroprevalence of Toxocara canis antibodies among Orang Asli (aborigines) in Peninsular Malaysia

Hakim SL, Mak JW, Lam PL, Nazma S, Normaznah Y

Southeast Asian J. Trop. Med. Public Health, 1992 Sep;23(3):493-6.
PMID: 1488706

An enzyme-linked immunosorbent assay using excretory-secretory antigens of the second stage larvae maintained in vitro was used to determine the seroprevalence of Toxocara antibodies in Orang Asli (aborigines) of Peninsular Malaysia. The mean + 3 SD optical density of 30 healthy subjects was used as the cut-off point. Overall prevalence was found to be 31.9%. No significant relationship was found between positive rates with sex and age groups, though children between 0 to 9 years recorded the highest positive rates. Eosinophil counts were found to be closely related to the proportion of positivity to toxocaral infection and mean optical densities. There was some degree of cross-reaction with Trichuris trichuria positive sera.

Matched MeSH terms: Antibodies, Helminth/blood*
Fulltext ELISA seropositivity for Toxocara canis antibodies in Malaysia, 1989-1991

Hakim SL, Mak JW, Lam PL

Med J Malaysia, 1993 Sep;48(3):303-7.
PMID: 8183143

The laboratory test results for visceral larva migrans (VLM) using ELISA for Toxocara canis antibodies employed by the Institute for Medical Research, Kuala Lumpur, is described. A total of 331 serum samples received from hospitals and general practitioners all over Malaysia were tested. The test utilises excretory-secretory antigens obtained from in vitro culture of second stage T. canis larvae. The overall seropositivity rate was 19.6%, the highest positive rate being in Indians (35.5%), followed by the Malays (14.8%), Chinese (10.9%) and others (29.4%). Seropositivity rate was highest in children below the age of 10; 89% of patients presented with eosinophilia and 93% with VLM syndrome were children.

Matched MeSH terms: Antibodies, Helminth/blood*
Prevalence of Toxocara canis antibody among children with bronchial asthma in Klang Hospital, Malaysia

Hakim SL, Thadasavanth M, Shamilah RH, Yogeswari S

Trans R Soc Trop Med Hyg, 1998 2 17;91(5):528.
PMID: 9463657 DOI: 10.1016/s0035-9203(97)90010-9
Matched MeSH terms: Antibodies, Helminth/blood
Recombinant expression of the larval excretory-secretory antigen TES-120 of Toxocara canis in the methylotrophic yeast Pichia pastoris

Fong MY, Lau YL

Parasitol Res, 2004 Jan;92(2):173-6.
PMID: 14655048

A gene encoding the larval excretory-secretory antigen TES-120 of the dog ascarid worm Toxocara canis was cloned into the methylotrophic yeast Pichia pastoris. Specificity of the recombinant TES-120 antigen produced by the yeast was investigated. Forty-five human serum samples from patients infected with different()parasitic organisms, including 8 cases of toxocariasis, were tested against the recombinant antigen in immunoblot assays. Results from the assays showed that the recombinant TES-120 antigen reacted with sera from toxocariasis patients only. This highly specific recombinant TES-120 antigen can potentially be used for the development of an inexpensive serodiagnostic assay for human toxocariasis.

Matched MeSH terms: Antibodies, Helminth/blood*
Fulltext Evaluation of dot immunogold filtration assay (DIGFA) for rapid serodiagnosis of eosinophilic meningitis due to Angio-strongylus cantonensis (Nematoda: Metastrongyloidea)

Eamsobhana P, Prasartvit A, Gan XX, Yong HS

Trop Biomed, 2015 Mar;32(1):121-5.
PMID: 25801261

Angiostrongylus cantonensis is the most frequent cause of eosinophilic meningitis in humans in Thailand and worldwide. Because of difficulty of recovering the Angiostrongylus larvae from infected patients, detection of parasite-specific antibodies is used to support clinical diagnosis. This study tested serum samples from eosinophilic meningitis patients and individuals at risk of infection with A. cantonensis to evaluate a recently developed simple and rapid dot-immunogold filtration assay (DIGFA) for detection of specific antibodies against A. cantonensis. Purified 31-kDa glycoprotein of A. cantonensis and protein A colloidal gold conjugate were employed to detect the 31-kDa anti-A. cantonensis antibody in patients sera from the parasite endemic areas of northeast Thailand. The results were compared with those obtained by dot-blot enzyme-linked immunosorbent assay (ELISA) with 31-kDa A. cantonensis antigen. The overall positivity rate of DIGFA and dot-blot ELISA for A. cantonensis infection in 98 clinically diagnosed cases from three highly endemic districts in Khon Kaen province were 39.79% and 37.75%, respectively. Among 86 sera of subjects at risk of infection with A. cantonensis, 24.41% were positive by DIGFA and 23.25% by dot-blot ELISA. There were good correlation between the visual grading of DIGFA and dot-blot ELISA in both groups of defined sera. DIGFA is as sensitive and specific as dot-blot ELISA for confirming eosinophilic meningitis due to A. cantonensis infection, with advantages of simplicity, rapidity and without the use of specific and expensive equipment, and can be used in field settings.

Matched MeSH terms: Antibodies, Helminth/blood*
Fulltext Immunochromatographic test for rapid serological diagnosis of human angiostrongyliasis

Eamsobhana P, Tungtrongchitr A, Wanachiwanawin D, Yong HS

Int J Infect Dis, 2018 Aug;73:69-71.
PMID: 29908250 DOI: 10.1016/j.ijid.2018.06.005

OBJECTIVES: The serological diagnosis of human infection with Angiostrongylus cantonensis remains problematic because there are no commercially available validated tests. Most laboratories use domestically prepared tests such as the enzyme-linked immunosorbent assay (ELISA) or immunoblotting. Since laboratory facilities are not always available in endemic areas, we developed and assessed a rapid lateral flow immunochromatographic assay (AcQuickDx Test) to detect anti-A. cantonensis antibodies in human serum.
METHODS: The test device was assembled with purified 31-kDa glycoprotein as diagnostic antigen and with gold-labelled anti-human immunoglublin-G as the detector reagent. A total of 97 serum samples were tested - 19 samples from clinically diagnosed patients with detectable A. cantonensis-specific antibody in immunoblotting; 43 samples from patients with other parasitic diseases, i.e. gnathostomiasis (n=13), toxocariasis (n=2), trichinellosis (n=2), hookworm infection (n=4), filariasis (n=5), cysticercosis (n=9), paragonimiasis (n=2), opisthorchiasis (n=3), and malaria (n=3); and 35 samples from normal healthy subjects.
RESULTS: The sensitivity, specificity, positive predictive value and negative predictive value of AcQuickDx Test to detect anti-A. cantonensis specific antibodies in serologically confirmed angiostrongyliasis cases, were 100%, 98.72%, 95% and 100%, respectively. Positive AcQuickDx was observed in 1 of 4 cases with hookworm infections. No positive AcQuickDx was observed in cases with other parasitic diseases, and the individual healthy subjects.
CONCLUSIONS: AcQuickDx Test is rapid, highly sensitive and specific, and easy to perform without additional equipment or ancillary supplies. It yields results that are interpreted visually, and possesses a long shelf-life at room temperature. Thus, it can be applied as an additional test for clinical diagnostic support of angiostrongyliasis either in conventional laboratories or for remote areas where laboratory infrastructure is not available.

Matched MeSH terms: Antibodies, Helminth/blood
Fulltext Serodiagnosis and early detection of Strongyloides stercoralis infection

Arifin N, Hanafiah KM, Ahmad H, Noordin R

J Microbiol Immunol Infect, 2019 Jun;52(3):371-378.
PMID: 30482708 DOI: 10.1016/j.jmii.2018.10.001

Strongyloidiasis is a major neglected tropical disease with the potential of causing lifelong infection and mortality. One of the ways for effective control of this disease is developing improved diagnostics, particularly using serological approaches. A serological test can achieve high diagnostic sensitivity and specificity, has the potential for point-of-care translation, and can be used as a screening tool for early detection. More research is needed to find clinically important antibody biomarkers for early disease detection, mapping, and epidemiological surveillance. This article summarizes human strongyloidiasis and the available diagnostic tools for the disease, focusing on describing the current antibody assays for strongyloidiasis. Finally, prospects of developing a more effective serodiagnostic tool for strongyloidiasis are discussed.

Matched MeSH terms: Antibodies, Helminth/blood*

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