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Species-Specific Serological Detection for Schistosomiasis by Serine Protease Inhibitor (SERPIN) in Multiplex Assay.

Tanigawa C, Fujii Y, Miura M, Nzou SM, Mwangi AW, Nagi S, Hamano S, Njenga SM, Mbanefo EC, Hirayama K, Mwau M, Kaneko S - PLoS Negl Trop Dis (2015)

Bottom Line: S. mansoni serine protease inhibitor (SERPIN) and Sm-RP26 showed significantly higher reactivity to patient plasma as compared to the control group.Sm-Filamin, Sm-GAPDH, Sm-GST, Sm-LAP1, Sm-LAP2, Sm-Sm31, Sm-Sm32 and Sm-Tropomyosin did not show difference in reactivity between S. mansoni infected and uninfected pupils.SERPINs showed correlation with the number of excreted eggs.

View Article: PubMed Central - PubMed

Affiliation: Department of Eco-Epidemiology, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan.

ABSTRACT

Background: Both Schistosoma mansoni and Schistosoma haematobium cause schistosomiasis in sub-Saharan Africa. We assessed the diagnostic value of selected Schistosoma antigens for the development of a multiplex serological immunoassay for sero-epidemiological surveillance.

Methodology/principal findings: Diagnostic ability of recombinant antigens from S. mansoni and S. haematobium was assessed by Luminex multiplex immunoassay using plasma from school children in two areas of Kenya, endemic for different species of schistosomiasis. S. mansoni serine protease inhibitor (SERPIN) and Sm-RP26 showed significantly higher reactivity to patient plasma as compared to the control group. Sm-Filamin, Sm-GAPDH, Sm-GST, Sm-LAP1, Sm-LAP2, Sm-Sm31, Sm-Sm32 and Sm-Tropomyosin did not show difference in reactivity between S. mansoni infected and uninfected pupils. Sm-RP26 was cross-reactive to plasma from S. haematobium patients, whereas Sm-SERPIN was species-specific. Sh-SEPRIN was partially cross-reactive to S. mansoni infected patients. ROC analysis for Sm-RP26, Sm-SERPIN and Sh-SERPIN showed AUC values of 0.833, 0.888 and 0.947, respectively. Using Spearman's rank correlation coefficient analysis, we also found significant positive correlation between the number of excreted eggs and median fluorescence intensity (MFI) from the multiplex immunoassays for Sm-SERPIN (ρ = 0.430, p-value = 0.003) and Sh-SERPIN (ρ = 0.433, p-value = 0.006).

Conclusions/significance: Sm-SERPIN is a promising species-specific diagnostic antigen. Sh-SEPRIN was partially cross-reactive to S. mansoni infected patients. SERPINs showed correlation with the number of excreted eggs. These indicate prospects for inclusion of SERPINs in the multiplex serological immunoassay system.

No MeSH data available.


Related in: MedlinePlus

Reactivity of Schistosoma mansoni recombinant antigens in the multiplex immunoassay.Multiplex assays were performed using samples from S. mansoni (Mbita) and S. haematobium (Kwale) endemic areas, respectively. Negative control samples from each location were included as comparator for the diagnostic efficacy of each antigen for the two species of schistosomes. The bar lines represent the median while the error bars represent the interquartile range. Antigens used in this assay: (A) Filamin; (B) GAPDH; (C) GST; (D) LAP-1; (E) LAP-2; (F) RP26; (G) Sm-SERPIN; (H) Sm31; (I) Sm32; (J) Tropomyosin-2; (K) SEA. Statistical significance was set at p < 0.05 as depicted using asterisks: * = p < 0.05, ** = p < 0.01, *** = p < 0.001, and **** = p < 0.0001. Jpn: healthy Japanese (n = 7), Sm-: egg negative in Sm endemic area (n = 26), Sm+: S. mansoni egg positive (n = 26), Sh-: egg negative in Sh endemic area (n = 26), Sh+: S. haematobium egg positive (n = 26).
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pntd.0004021.g003: Reactivity of Schistosoma mansoni recombinant antigens in the multiplex immunoassay.Multiplex assays were performed using samples from S. mansoni (Mbita) and S. haematobium (Kwale) endemic areas, respectively. Negative control samples from each location were included as comparator for the diagnostic efficacy of each antigen for the two species of schistosomes. The bar lines represent the median while the error bars represent the interquartile range. Antigens used in this assay: (A) Filamin; (B) GAPDH; (C) GST; (D) LAP-1; (E) LAP-2; (F) RP26; (G) Sm-SERPIN; (H) Sm31; (I) Sm32; (J) Tropomyosin-2; (K) SEA. Statistical significance was set at p < 0.05 as depicted using asterisks: * = p < 0.05, ** = p < 0.01, *** = p < 0.001, and **** = p < 0.0001. Jpn: healthy Japanese (n = 7), Sm-: egg negative in Sm endemic area (n = 26), Sm+: S. mansoni egg positive (n = 26), Sh-: egg negative in Sh endemic area (n = 26), Sh+: S. haematobium egg positive (n = 26).

Mentions: Only RP26 and Sm-SERPIN were specifically reactive to plasma from S. mansoni patients (Fig 3). The other antigens did not show disease-specific reactivity, although the reactivity to plasma from endemic areas is higher than reactivity to Japanese control as would be expected. The two antigens with disease-specific reactivity (Sm-RP26 and Sm-SERPIN) showed fluorescence signals (MFI) that are similar or higher than signals recorded from evaluation of coupling with anti-polyhistidine-tag antibody (Fig 2A). This further shows that the absence of reactivity from the other 8 antigens was not a function of number of coupled antigens. For instance, Sm-RP26 antigen with relatively low microsphere-coupled antigens based on evaluation with anti-polyhistidine-tag antibody showed significant disease-specific reactivity. Interestingly, Sm-RP26 was also reactive to plasma from S. haematobium patients (Fig 3). Our data also showed that Sm-SERPIN was efficient in species-specific differential detection of infections with S. mansoni. Also, S. haematobium SERPIN was previously reported to be species-specific [26]. The crude egg antigen (SEA) from S. mansoni showed similar MFI value between egg negative and positive in Mbita area, where S. mansoni is endemic; suggesting that some egg negative individuals may have history of previous infections. Surprisingly, SEA from S. mansoni showed disease-specific detection only for S. haematobium patients (Fig 3). Taken together, our data showed that Sm-SERPIN and Sm-RP26 are promising diagnostic candidates to be potentially included in our multiplex system. In addition, Sm-SERPIN demonstrated species-specific reactivity.


Species-Specific Serological Detection for Schistosomiasis by Serine Protease Inhibitor (SERPIN) in Multiplex Assay.

Tanigawa C, Fujii Y, Miura M, Nzou SM, Mwangi AW, Nagi S, Hamano S, Njenga SM, Mbanefo EC, Hirayama K, Mwau M, Kaneko S - PLoS Negl Trop Dis (2015)

Reactivity of Schistosoma mansoni recombinant antigens in the multiplex immunoassay.Multiplex assays were performed using samples from S. mansoni (Mbita) and S. haematobium (Kwale) endemic areas, respectively. Negative control samples from each location were included as comparator for the diagnostic efficacy of each antigen for the two species of schistosomes. The bar lines represent the median while the error bars represent the interquartile range. Antigens used in this assay: (A) Filamin; (B) GAPDH; (C) GST; (D) LAP-1; (E) LAP-2; (F) RP26; (G) Sm-SERPIN; (H) Sm31; (I) Sm32; (J) Tropomyosin-2; (K) SEA. Statistical significance was set at p < 0.05 as depicted using asterisks: * = p < 0.05, ** = p < 0.01, *** = p < 0.001, and **** = p < 0.0001. Jpn: healthy Japanese (n = 7), Sm-: egg negative in Sm endemic area (n = 26), Sm+: S. mansoni egg positive (n = 26), Sh-: egg negative in Sh endemic area (n = 26), Sh+: S. haematobium egg positive (n = 26).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4546333&req=5

pntd.0004021.g003: Reactivity of Schistosoma mansoni recombinant antigens in the multiplex immunoassay.Multiplex assays were performed using samples from S. mansoni (Mbita) and S. haematobium (Kwale) endemic areas, respectively. Negative control samples from each location were included as comparator for the diagnostic efficacy of each antigen for the two species of schistosomes. The bar lines represent the median while the error bars represent the interquartile range. Antigens used in this assay: (A) Filamin; (B) GAPDH; (C) GST; (D) LAP-1; (E) LAP-2; (F) RP26; (G) Sm-SERPIN; (H) Sm31; (I) Sm32; (J) Tropomyosin-2; (K) SEA. Statistical significance was set at p < 0.05 as depicted using asterisks: * = p < 0.05, ** = p < 0.01, *** = p < 0.001, and **** = p < 0.0001. Jpn: healthy Japanese (n = 7), Sm-: egg negative in Sm endemic area (n = 26), Sm+: S. mansoni egg positive (n = 26), Sh-: egg negative in Sh endemic area (n = 26), Sh+: S. haematobium egg positive (n = 26).
Mentions: Only RP26 and Sm-SERPIN were specifically reactive to plasma from S. mansoni patients (Fig 3). The other antigens did not show disease-specific reactivity, although the reactivity to plasma from endemic areas is higher than reactivity to Japanese control as would be expected. The two antigens with disease-specific reactivity (Sm-RP26 and Sm-SERPIN) showed fluorescence signals (MFI) that are similar or higher than signals recorded from evaluation of coupling with anti-polyhistidine-tag antibody (Fig 2A). This further shows that the absence of reactivity from the other 8 antigens was not a function of number of coupled antigens. For instance, Sm-RP26 antigen with relatively low microsphere-coupled antigens based on evaluation with anti-polyhistidine-tag antibody showed significant disease-specific reactivity. Interestingly, Sm-RP26 was also reactive to plasma from S. haematobium patients (Fig 3). Our data also showed that Sm-SERPIN was efficient in species-specific differential detection of infections with S. mansoni. Also, S. haematobium SERPIN was previously reported to be species-specific [26]. The crude egg antigen (SEA) from S. mansoni showed similar MFI value between egg negative and positive in Mbita area, where S. mansoni is endemic; suggesting that some egg negative individuals may have history of previous infections. Surprisingly, SEA from S. mansoni showed disease-specific detection only for S. haematobium patients (Fig 3). Taken together, our data showed that Sm-SERPIN and Sm-RP26 are promising diagnostic candidates to be potentially included in our multiplex system. In addition, Sm-SERPIN demonstrated species-specific reactivity.

Bottom Line: S. mansoni serine protease inhibitor (SERPIN) and Sm-RP26 showed significantly higher reactivity to patient plasma as compared to the control group.Sm-Filamin, Sm-GAPDH, Sm-GST, Sm-LAP1, Sm-LAP2, Sm-Sm31, Sm-Sm32 and Sm-Tropomyosin did not show difference in reactivity between S. mansoni infected and uninfected pupils.SERPINs showed correlation with the number of excreted eggs.

View Article: PubMed Central - PubMed

Affiliation: Department of Eco-Epidemiology, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan.

ABSTRACT

Background: Both Schistosoma mansoni and Schistosoma haematobium cause schistosomiasis in sub-Saharan Africa. We assessed the diagnostic value of selected Schistosoma antigens for the development of a multiplex serological immunoassay for sero-epidemiological surveillance.

Methodology/principal findings: Diagnostic ability of recombinant antigens from S. mansoni and S. haematobium was assessed by Luminex multiplex immunoassay using plasma from school children in two areas of Kenya, endemic for different species of schistosomiasis. S. mansoni serine protease inhibitor (SERPIN) and Sm-RP26 showed significantly higher reactivity to patient plasma as compared to the control group. Sm-Filamin, Sm-GAPDH, Sm-GST, Sm-LAP1, Sm-LAP2, Sm-Sm31, Sm-Sm32 and Sm-Tropomyosin did not show difference in reactivity between S. mansoni infected and uninfected pupils. Sm-RP26 was cross-reactive to plasma from S. haematobium patients, whereas Sm-SERPIN was species-specific. Sh-SEPRIN was partially cross-reactive to S. mansoni infected patients. ROC analysis for Sm-RP26, Sm-SERPIN and Sh-SERPIN showed AUC values of 0.833, 0.888 and 0.947, respectively. Using Spearman's rank correlation coefficient analysis, we also found significant positive correlation between the number of excreted eggs and median fluorescence intensity (MFI) from the multiplex immunoassays for Sm-SERPIN (ρ = 0.430, p-value = 0.003) and Sh-SERPIN (ρ = 0.433, p-value = 0.006).

Conclusions/significance: Sm-SERPIN is a promising species-specific diagnostic antigen. Sh-SEPRIN was partially cross-reactive to S. mansoni infected patients. SERPINs showed correlation with the number of excreted eggs. These indicate prospects for inclusion of SERPINs in the multiplex serological immunoassay system.

No MeSH data available.


Related in: MedlinePlus