1.Monoclonal Antibody-Based Dipstick Assay: A Reliable Field Applicable Technique for Diagnosis of Schistosoma mansoni Infection Using Human Serum and Urine Samples.
Zeinab DEMERDASH ; Salwa MOHAMED ; Mohamed HENDAWY ; Ibrahim RABIA ; Mohy ATTIA ; Zeinab SHAKER ; Tarek M DIAB
The Korean Journal of Parasitology 2013;51(1):93-98
A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.
Animals
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Antibodies, Helminth/diagnostic use/isolation & purification
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Antibodies, Monoclonal/diagnostic use/isolation & purification
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Antigens, Helminth/*blood/*urine
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Diagnostic Tests, Routine/*methods
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Humans
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Immunoassay/methods
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Parasitology/*methods
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*Point-of-Care Systems
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Schistosoma mansoni/immunology/*isolation & purification
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Schistosomiasis mansoni/*diagnosis
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Sensitivity and Specificity
2.A Recombinant Matrix Metalloproteinase Protein from Gnathostoma spinigerum for Serodiagnosis of Neurognathostomiasis.
Penchom JANWAN ; Pewpan M INTAPAN ; Hiroshi YAMASAKI ; Porntip LAUMMAUNWAI ; Kittisak SAWANYAWISUTH ; Chaisiri WONGKHAM ; Chatchai TAYAPIWATANA ; Amnat KITKHUANDEE ; Viraphong LULITANOND ; Yukifumi NAWA ; Wanchai MALEEWONG
The Korean Journal of Parasitology 2013;51(6):751-754
Neurognathostomiasis is a severe form of human gnathostomiasis which can lead to disease and death. Diagnosis of neurognathostomiasis is made presumptively by using clinical manifestations. Immunoblotting, which recognizes antigenic components of molecular mass 21 kDa and 24 kDa in larval extracts of Gnathostoma spinigerum (Gs 21/24), has high sensitivity and specificity for diagnosis of neurognathostomiasis. However, only very small amounts of the Gs 21/24 antigens can be prepared from parasites harvested from natural or experimental animals. To overcome this problem, we recently produced a recombinant matrix metalloproteinase (rMMP) protein from G. spinigerum. In this study, we evaluated this rMMP alongside the Gs 21/24 antigens for serodiagnosis of human neurognathostomiasis. We studied sera from 40 patients from Srinagarind Hospital, Khon Kaen University, Thailand, with clinical criteria consistent with those of neurognathostomiasis, and sera from 30 healthy control adults from Thailand. All sera were tested for specific IgG antibodies against both G. spinigerum crude larval extract and rMMP protein using immunoblot analysis. The sensitivity and specificity for both antigenic preparations were all 100%. These results show that G. spinigerum rMMP protein can be used as an alternative diagnostic antigen, in place of larval extract, for serodiagnosis of neurognathostomiasis.
Adult
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Animals
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Antibodies, Helminth/blood
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Antigens, Helminth/*diagnostic use/genetics/isolation & purification
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Central Nervous System Parasitic Infections/*diagnosis/parasitology
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Gnathostoma/enzymology/immunology/*isolation & purification
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Gnathostomiasis/*diagnosis/parasitology
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Healthy Volunteers
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Humans
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Immunoblotting/methods
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Immunoglobulin G/blood
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Matrix Metalloproteinases/*diagnostic use/genetics/isolation & purification
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Parasitology/*methods
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Prospective Studies
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Recombinant Proteins/diagnostic use/genetics/isolation & purification
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Sensitivity and Specificity
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Serologic Tests/methods
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Thailand
3.Dot-Blot Immunoassay of Fasciola gigantica Infection using 27 kDa and Adult Worm Regurge Antigens in Egyptian Patients.
Hanan H KAMEL ; Ghada A SAAD ; Rania M SARHAN
The Korean Journal of Parasitology 2013;51(2):177-182
The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27-(KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.
Animals
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Antibodies, Helminth/*blood
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Antigens, Helminth/*diagnostic use
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Diagnostic Tests, Routine/*methods
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Egypt
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Fasciola/immunology/*isolation & purification
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Fascioliasis/*diagnosis/parasitology
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Humans
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Immunoblotting/*methods
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Parasitology/*methods
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Predictive Value of Tests
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Sensitivity and Specificity
4.Detection of Gnathostoma spinigerum Antibodies in Sera of Non-Traumatic Subarachnoid Hemorrhage Patients in Thailand.
Amnat KITKHUANDEE ; Waranon MUNKONG ; Kittisak SAWANYAWISUTH ; Penchom JANWAN ; Wanchai MALEEWONG ; Pewpan M INTAPAN
The Korean Journal of Parasitology 2013;51(6):755-757
Gnathostoma spinigerum can cause subarachnoid hemorrhage (SAH). The detection of specific antibodies in serum against G. spinigerum antigen is helpful for diagnosis of neurognathostomiasis. There is limited data on the frequency of G. spinigerum infection in non-traumatic SAH. A series of patients diagnosed as non-traumatic SAH at the Srinagarind Hospital, Khon Kaen University, Thailand between January 2011 and January 2013 were studied. CT or MR imaging of the brain was used for diagnosis of SAH. Patients were categorized as aneurysmal subarachnoid hemorrhage (A-SAH) or non-aneurysmal subarachnoid hemorrhage (NA-SAH) according to the results of cerebral angiograms. The presence of specific antibodies in serum against 21- or 24-kDa G. spinigerum antigen was determined using the immunoblot technique. The detection rate of antibodies was compared between the 2 groups. Of the 118 non-traumatic SAH patients for whom cerebral angiogram and immunoblot data were available, 80 (67.8%) patients had A-SAH, whereas 38 (32.2%) had NA-SAH. Overall, 23.7% were positive for specific antibodies against 21- and/or 24-kDa G. spinigerum antigen. No significant differences were found in the positive rate of specific antibodies against G. spinigerum in both groups (P-value=0.350).
Adult
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Aged
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Animals
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Antibodies, Helminth/*blood
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Antigens, Helminth/diagnostic use
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Brain/radiography
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Female
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Gnathostoma/immunology/*isolation & purification
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Gnathostomiasis/*diagnosis/*parasitology
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Humans
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Immunoblotting
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Magnetic Resonance Imaging
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Male
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Middle Aged
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Serum/immunology
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Subarachnoid Hemorrhage/*diagnosis/*etiology
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Thailand
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Tomography, X-Ray Computed