Evaluation of combined B cell specific N-terminal immunogenic domains of LipL21 for diagnosis of leptospirosis

Kumari Anita, Mallela Martha Premlatha, Murugesan Kanagavel, Charles Solomon Akino Mercy, Veerapandian Raja, Santhanam Shanmughapriya, Kalimuthusamy Natarajaseenivasan

Research output: Contribution to journalArticle

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Abstract

Leptospiral outer membrane protein LipL21 and its truncated N-terminal immunogenic region (I-LipL21) were evaluated for diagnosis of leptospirosis. The complete coding sequence of LipL21 nucleotide sequence was subjected to BCPred and VaxiJen analysis for determination of B cell specific immunogenic epitopes. Epitope1 ACS STD TGQ KDA TTV GDG (1.8837), Epitope2 WGG PPE QRN DGK TPR DTN (0.9483), Epitope3 VKG VGV YEC KAT GSG SDP (1.4077) and Epitope4 NEW ECQ CVI YAK FPG GKD (0.4462) were predicted. LipL21 and N-terminal fragment having B-cell specific epitopes with higher VaxiJen score >0.9 as truncated I-LipL21 were cloned independently in pET15b and expressed in Escherichia coli. IgM ELISA and dot blot assay was performed for sera samples collected from Delhi-NCR for leptospiral whole cell lysate (WCL), recombinant LipL21 and I-LipL21. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were found to be 92.5%, 92.8%, 83.3%, and 97% respectively for recombinant I-LipL21 by IgM-ELISA. 11-14.8% increased sensitivity was observed over LipL21 and WCL. The I-LipL21 dot blot assay showed a further increased sensitivity of 3.8% over the IgM-ELISA. Therefore I-LipL21 may be the ideal candidate protein for diagnosis of leptospirosis.

Original languageEnglish (US)
Pages (from-to)465-470
Number of pages6
JournalInternational Journal of Biological Macromolecules
Volume91
DOIs
StatePublished - Oct 1 2016

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Epitopes
Leptospirosis
Immunoglobulin M
Assays
B-Lymphocytes
Enzyme-Linked Immunosorbent Assay
Cells
Proteins
Nucleotides
Escherichia coli
B-Lymphocyte Epitopes
Sexually Transmitted Diseases
Membranes
Membrane Proteins
Sensitivity and Specificity
Serum
Evaluation
Enzyme-linked immunosorbent assay
Protein
Recombinant

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Economics and Econometrics
  • Energy(all)

Cite this

Anita, Kumari ; Premlatha, Mallela Martha ; Kanagavel, Murugesan ; Akino Mercy, Charles Solomon ; Raja, Veerapandian ; Shanmughapriya, Santhanam ; Natarajaseenivasan, Kalimuthusamy. / Evaluation of combined B cell specific N-terminal immunogenic domains of LipL21 for diagnosis of leptospirosis. In: International Journal of Biological Macromolecules. 2016 ; Vol. 91. pp. 465-470.
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abstract = "Leptospiral outer membrane protein LipL21 and its truncated N-terminal immunogenic region (I-LipL21) were evaluated for diagnosis of leptospirosis. The complete coding sequence of LipL21 nucleotide sequence was subjected to BCPred and VaxiJen analysis for determination of B cell specific immunogenic epitopes. Epitope1 ACS STD TGQ KDA TTV GDG (1.8837), Epitope2 WGG PPE QRN DGK TPR DTN (0.9483), Epitope3 VKG VGV YEC KAT GSG SDP (1.4077) and Epitope4 NEW ECQ CVI YAK FPG GKD (0.4462) were predicted. LipL21 and N-terminal fragment having B-cell specific epitopes with higher VaxiJen score >0.9 as truncated I-LipL21 were cloned independently in pET15b and expressed in Escherichia coli. IgM ELISA and dot blot assay was performed for sera samples collected from Delhi-NCR for leptospiral whole cell lysate (WCL), recombinant LipL21 and I-LipL21. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were found to be 92.5{\%}, 92.8{\%}, 83.3{\%}, and 97{\%} respectively for recombinant I-LipL21 by IgM-ELISA. 11-14.8{\%} increased sensitivity was observed over LipL21 and WCL. The I-LipL21 dot blot assay showed a further increased sensitivity of 3.8{\%} over the IgM-ELISA. Therefore I-LipL21 may be the ideal candidate protein for diagnosis of leptospirosis.",
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Evaluation of combined B cell specific N-terminal immunogenic domains of LipL21 for diagnosis of leptospirosis. / Anita, Kumari; Premlatha, Mallela Martha; Kanagavel, Murugesan; Akino Mercy, Charles Solomon; Raja, Veerapandian; Shanmughapriya, Santhanam; Natarajaseenivasan, Kalimuthusamy.

In: International Journal of Biological Macromolecules, Vol. 91, 01.10.2016, p. 465-470.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of combined B cell specific N-terminal immunogenic domains of LipL21 for diagnosis of leptospirosis

AU - Anita, Kumari

AU - Premlatha, Mallela Martha

AU - Kanagavel, Murugesan

AU - Akino Mercy, Charles Solomon

AU - Raja, Veerapandian

AU - Shanmughapriya, Santhanam

AU - Natarajaseenivasan, Kalimuthusamy

PY - 2016/10/1

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N2 - Leptospiral outer membrane protein LipL21 and its truncated N-terminal immunogenic region (I-LipL21) were evaluated for diagnosis of leptospirosis. The complete coding sequence of LipL21 nucleotide sequence was subjected to BCPred and VaxiJen analysis for determination of B cell specific immunogenic epitopes. Epitope1 ACS STD TGQ KDA TTV GDG (1.8837), Epitope2 WGG PPE QRN DGK TPR DTN (0.9483), Epitope3 VKG VGV YEC KAT GSG SDP (1.4077) and Epitope4 NEW ECQ CVI YAK FPG GKD (0.4462) were predicted. LipL21 and N-terminal fragment having B-cell specific epitopes with higher VaxiJen score >0.9 as truncated I-LipL21 were cloned independently in pET15b and expressed in Escherichia coli. IgM ELISA and dot blot assay was performed for sera samples collected from Delhi-NCR for leptospiral whole cell lysate (WCL), recombinant LipL21 and I-LipL21. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were found to be 92.5%, 92.8%, 83.3%, and 97% respectively for recombinant I-LipL21 by IgM-ELISA. 11-14.8% increased sensitivity was observed over LipL21 and WCL. The I-LipL21 dot blot assay showed a further increased sensitivity of 3.8% over the IgM-ELISA. Therefore I-LipL21 may be the ideal candidate protein for diagnosis of leptospirosis.

AB - Leptospiral outer membrane protein LipL21 and its truncated N-terminal immunogenic region (I-LipL21) were evaluated for diagnosis of leptospirosis. The complete coding sequence of LipL21 nucleotide sequence was subjected to BCPred and VaxiJen analysis for determination of B cell specific immunogenic epitopes. Epitope1 ACS STD TGQ KDA TTV GDG (1.8837), Epitope2 WGG PPE QRN DGK TPR DTN (0.9483), Epitope3 VKG VGV YEC KAT GSG SDP (1.4077) and Epitope4 NEW ECQ CVI YAK FPG GKD (0.4462) were predicted. LipL21 and N-terminal fragment having B-cell specific epitopes with higher VaxiJen score >0.9 as truncated I-LipL21 were cloned independently in pET15b and expressed in Escherichia coli. IgM ELISA and dot blot assay was performed for sera samples collected from Delhi-NCR for leptospiral whole cell lysate (WCL), recombinant LipL21 and I-LipL21. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were found to be 92.5%, 92.8%, 83.3%, and 97% respectively for recombinant I-LipL21 by IgM-ELISA. 11-14.8% increased sensitivity was observed over LipL21 and WCL. The I-LipL21 dot blot assay showed a further increased sensitivity of 3.8% over the IgM-ELISA. Therefore I-LipL21 may be the ideal candidate protein for diagnosis of leptospirosis.

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