Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

Arora, R.; Singh, Nisha; Juyal, P. D.; Kumari, Jyoti; Ghosh, Srikant

doi:10.1007/s12639-010-0011-6

Cited by 10 publications

(7 citation statements)

References 21 publications

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“…No polypeptide band showed immunoreactivity with the verified pooled negative serum Previous studies on Western blot analysis of somatic antigens of G. crumenifer (Saifullah et al, 2000;Saifullah et al, 2011), recorded low molecular weight polypeptides, ranging from <14 to 50 kDa. The results of studies on other trematodes are comparable with the current research, having identified somatic antigenic polypeptides of: F. hepatica in the range of 33 and 66 kDa (Gonenc et al, 2004); Paramphistomum cervi 52 kDa (Anuracpreeda et al, 2008); mixed infection with paramphistomes 90 kDa (Meshgi et al, 2009); and P. epiclitum with two lower molecular weight protein bands of 37.6 and 39.8 kDa (Arora et al, 2010). The studies on Paramphistomum spp.…”

Section: Discussionsupporting

confidence: 84%

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Serological diagnostic potential of the 38-72 kDa somatic antigen of Gastrothylax crumenifer in buffaloes using the Indirect Enzyme-Linked Immunosorbent Assay

Ali

Afshan

Fuentes

et al. 2022

J Hellenic Vet Med Soc

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Paramphistomosis, caused by digenetic trematodes of the superfamily Paramphistomoidea, causes heavy economic losses in terms of reduced fertility, milk and meat production in the livestock industry. In the present study, somatic and excretory secretory antigens isolated from 500 live Gastrothylax crumenifer were assessed for their diagnostic potential by using an antibody detection enzyme immunoassay. Prior to the enzyme immunoassay, the somatic and excretory/secretory (ES) antigens of G. crumenifer were subjected to SDS-PAGE and Western blot (WB) for the detection of immunogenic proteins. Indirect ELISA analysis was performed on sera from buffaloes naturally infected with G. crumenifer, with control sera of buffaloes infected with Gigantocotyle explanatum, Fasciola spp., Cotylophoron /Paramphistomum spp. The SDS-PAGE results of the somatic products of G. crumenifer identified proteins were between 10-123 kDa, showing a maximum abundance of 10, 15, 25-28, 36, 38-72, 95-123 kDa proteins. The most abundant proteins recorded in ES product were ≥95, 72, 55 and 40 kDa. The antigenic analysis of somatic proteins using WB revealed reactive polypeptides of a size between 55-70 kDa, while metabolic extracts did not show reactivity with naturally infected buffalo sera. The sensitivity and specificity of the ELISA test for 38-72 kDa somatic antigens were 85.71% and 89.74%, respectively. The cross-reactivity with other trematode sera was 16-20%. Antibodies were tested against the 38-72 kDa somatic antigens, and 19.69% (39/198) of buffaloes were found positive, while 12.1% (24/198) presented infection in the fecal/postmortem examination. The study confirmed that ELISA established for the 38-72 kDa somatic antigens of G. crumenifer had diagnostic potential against paramphistome infections.

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Section: Discussionsupporting

confidence: 84%

“…Similarly, studies on P. epiclitum recorded molecular weight bands in the ranges of 4-100 kDa (Jadav et al, 2018) and 14.9-95.5 kDa (Arora et al, 2010); in P. cervi the molecular weight band ranged from 12 to 100 kDa (Dar et al, 2016), and in Paramphistomum spp. the molecular weight band was in the 11.5-174 kDa range (Salib et al, 2015).…”

Section: Discussionmentioning

confidence: 83%

Serological diagnostic potential of the 38-72 kDa somatic antigen of Gastrothylax crumenifer in buffaloes using the Indirect Enzyme-Linked Immunosorbent Assay

Ali

Afshan

Fuentes

et al. 2022

J Hellenic Vet Med Soc

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“…The existence of crossreaction that occurred in helminthiasis infection could overcome by the use of purified proteins. The use of serological diagnosis techniques on a large scale could be done by using serum obtained from farms, abattoir and experimental animals to determine the level of sensitivity and specificity of the test (Arora et al, 2010).…”

Section: Resultsmentioning

confidence: 99%

Western Blot Analysis to Detect Cross-reaction in Toxocara vitulorum Protein with Anti-Mecistocirrus digitatus Serum

Anggraini¹,

Kusnoto²,

Sarudji³

2021

WVJ

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Worm infections are found in livestock and can be transmitted to humans. Toxocara vitulorum is a worm species which commonly infected people. Cross-reaction among worms can generate false positive to establish helminthiasis diagnosis through antibody inspection. This study aimed to determine specific proteins that caused cross-reaction between Toxocara vitulorum antigen and anti-M. digitatus serum by using the western blot technique. In the present study, the whole worms extracted of T. vitulorum and M. digitatus have been used to make polyclonal antibodies from M. digitatus with Wistar rats as hosts. The cross-reaction between whole worm extract of T. vitulorum protein and anti-M. digitatus serum obtained 12 protein bands that each relative molecular mass (Mr) valued of 176, 124, 92, 68, 59, 47, 31, 29, 26, 16, 12, and 10 kDa. Cross-reaction occurred between T. vitulorum protein and anti-M. digitatus.

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“…The cattle and buffaloes postmortem examination showed positivity with other trematodes along with fasciolid infections and cross reactivity of 10 to 20% was observed for developed ELISA (Table 1). This may also explain with some common antigenic epitopes that are capable of detecting antigens from other trematodes (Arora et al, 2010).…”

Section: Elisamentioning

confidence: 93%

Diagnostic potential of 36-55 kDa somatic antigens of Fasciola gigantica for bovine fasciolosis

et al. 2022

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Fascioliasis causes huge economic losses in livestock industry by reducing the growth, fertility rate, meat and milk yield. In the present study, somatic antigens isolated from Fasciola gigantica were assessed for the early detection of infection by developing antibody detection enzyme immunoassay. The somatic antigens extracted from adult F. gigantica worms was separated on SDS-PAGE and immunogenicity was determined by Western blot (WB). The results showed polypeptides bands between 15 to 95 kDa, and most prominent bands were 15, 28, 36, 38, 55, 72 and 95 kDa polypeptides. The WB revealed a single polypeptide of size between 36 to 55 kDa as antigenic. The sensitivity and specificity of ELISA test established for 36 to 55 kDa somatic antigens was 95.45% (95% CI: 77.16% to 99.88%) and 87.1% (95% CI: 70.17% to 96.37%), respectively. Kappa value revealed that the strength of agreement is considered to be very ‘good’. In house established ELISA was implemented in the field and 12.1% (31/256) large ruminants were found positive, while 4.68% (12/256) with postmortem examination. The fasciolosis did not show significant (P>0.05) association with host type, breed and age groups, while significant (P<0.01) association with host sexes. The result confirmed that in-house established ELISA test had good value for serodiagnosis of fasciolosis in cattle and buffaloes for large scale epidemiological studies.

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Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum

Cited by 10 publications

References 21 publications

Serological diagnostic potential of the 38-72 kDa somatic antigen of Gastrothylax crumenifer in buffaloes using the Indirect Enzyme-Linked Immunosorbent Assay

Serological diagnostic potential of the 38-72 kDa somatic antigen of Gastrothylax crumenifer in buffaloes using the Indirect Enzyme-Linked Immunosorbent Assay

Western Blot Analysis to Detect Cross-reaction in Toxocara vitulorum Protein with Anti-Mecistocirrus digitatus Serum

Diagnostic potential of 36-55 kDa somatic antigens of Fasciola gigantica for bovine fasciolosis

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