In vivo-expressed proteins of virulent Leptospira interrogans serovar autumnalis N2 elicit strong IgM responses of value in conclusive diagnosis

Veerapandian Raja, Santhanam Shanmughapriya, Murugesan Kanagavel, Sergey C. Artiushin, Sridhar Velineni, John F. Timoney, Kalimuthusamy Natarajaseenivasan

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Leptospirosis is a serious zoonosis that is underdiagnosed because of limited access to laboratory facilities in Southeast Asia, Central and South America, and Oceania. Timely diagnosis of locally distributed serovars of high virulence is crucial for successful care and outbreak management. Using pooled patient sera, an expression gene library of a virulent Leptospira interrogans serovar Autumnalis strain N2 isolated in South India was screened. The identified genes were characterized, and the purified recombinant proteins were used as antigens in IgM enzyme-linked immunosorbent assay (ELISA) either singly or in combination. Sera (n=118) from cases of acute leptospirosis along with sera (n=58) from healthy subjects were tested for reactivity with the identified proteins in an ELISA designed to detect specific IgM responses. We have identified nine immunoreactive proteins, ArgC, RecA, GlpF, FliD, TrmD, RplS, RnhB, Lp28.6, and Lrr44.9, which were found to be highly conserved among pathogenic leptospires. Apparently, the proteins ArgC, RecA, GlpF, FliD, TrmD, and Lrr44.9 are expressed during natural infection of the host and undetectable in in vitro cultures. Among all the recombinant proteins used as antigens in IgM ELISA, ArgC had the highest sensitivity and specificity, 89.8% and 95.5%, respectively, for the conclusive diagnosis of leptospirosis. The use of ArgC and RecA in combination for IgM ELISA increased the sensitivity and specificity to 95.7% and 94.9%, respectively. ArgC and RecA thus elicited specific IgM responses and were therefore effective in laboratory confirmation of Leptospira infection.

Original languageEnglish (US)
Pages (from-to)65-72
Number of pages8
JournalClinical and Vaccine Immunology
Volume23
Issue number1
DOIs
StatePublished - Jan 2016

Fingerprint

Leptospira interrogans serovar autumnalis
Immunosorbents
Immunoglobulin M
Leptospirosis
Assays
Enzyme-Linked Immunosorbent Assay
Rec A Recombinases
Enzymes
Proteins
Recombinant Proteins
Serum
Oceania
Antigens
Central America
Leptospira
Sensitivity and Specificity
Southeastern Asia
South America
Zoonoses
Infection

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

Cite this

Raja, Veerapandian ; Shanmughapriya, Santhanam ; Kanagavel, Murugesan ; Artiushin, Sergey C. ; Velineni, Sridhar ; Timoney, John F. ; Natarajaseenivasan, Kalimuthusamy. / In vivo-expressed proteins of virulent Leptospira interrogans serovar autumnalis N2 elicit strong IgM responses of value in conclusive diagnosis. In: Clinical and Vaccine Immunology. 2016 ; Vol. 23, No. 1. pp. 65-72.
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abstract = "Leptospirosis is a serious zoonosis that is underdiagnosed because of limited access to laboratory facilities in Southeast Asia, Central and South America, and Oceania. Timely diagnosis of locally distributed serovars of high virulence is crucial for successful care and outbreak management. Using pooled patient sera, an expression gene library of a virulent Leptospira interrogans serovar Autumnalis strain N2 isolated in South India was screened. The identified genes were characterized, and the purified recombinant proteins were used as antigens in IgM enzyme-linked immunosorbent assay (ELISA) either singly or in combination. Sera (n=118) from cases of acute leptospirosis along with sera (n=58) from healthy subjects were tested for reactivity with the identified proteins in an ELISA designed to detect specific IgM responses. We have identified nine immunoreactive proteins, ArgC, RecA, GlpF, FliD, TrmD, RplS, RnhB, Lp28.6, and Lrr44.9, which were found to be highly conserved among pathogenic leptospires. Apparently, the proteins ArgC, RecA, GlpF, FliD, TrmD, and Lrr44.9 are expressed during natural infection of the host and undetectable in in vitro cultures. Among all the recombinant proteins used as antigens in IgM ELISA, ArgC had the highest sensitivity and specificity, 89.8{\%} and 95.5{\%}, respectively, for the conclusive diagnosis of leptospirosis. The use of ArgC and RecA in combination for IgM ELISA increased the sensitivity and specificity to 95.7{\%} and 94.9{\%}, respectively. ArgC and RecA thus elicited specific IgM responses and were therefore effective in laboratory confirmation of Leptospira infection.",
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In vivo-expressed proteins of virulent Leptospira interrogans serovar autumnalis N2 elicit strong IgM responses of value in conclusive diagnosis. / Raja, Veerapandian; Shanmughapriya, Santhanam; Kanagavel, Murugesan; Artiushin, Sergey C.; Velineni, Sridhar; Timoney, John F.; Natarajaseenivasan, Kalimuthusamy.

In: Clinical and Vaccine Immunology, Vol. 23, No. 1, 01.2016, p. 65-72.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vivo-expressed proteins of virulent Leptospira interrogans serovar autumnalis N2 elicit strong IgM responses of value in conclusive diagnosis

AU - Raja, Veerapandian

AU - Shanmughapriya, Santhanam

AU - Kanagavel, Murugesan

AU - Artiushin, Sergey C.

AU - Velineni, Sridhar

AU - Timoney, John F.

AU - Natarajaseenivasan, Kalimuthusamy

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N2 - Leptospirosis is a serious zoonosis that is underdiagnosed because of limited access to laboratory facilities in Southeast Asia, Central and South America, and Oceania. Timely diagnosis of locally distributed serovars of high virulence is crucial for successful care and outbreak management. Using pooled patient sera, an expression gene library of a virulent Leptospira interrogans serovar Autumnalis strain N2 isolated in South India was screened. The identified genes were characterized, and the purified recombinant proteins were used as antigens in IgM enzyme-linked immunosorbent assay (ELISA) either singly or in combination. Sera (n=118) from cases of acute leptospirosis along with sera (n=58) from healthy subjects were tested for reactivity with the identified proteins in an ELISA designed to detect specific IgM responses. We have identified nine immunoreactive proteins, ArgC, RecA, GlpF, FliD, TrmD, RplS, RnhB, Lp28.6, and Lrr44.9, which were found to be highly conserved among pathogenic leptospires. Apparently, the proteins ArgC, RecA, GlpF, FliD, TrmD, and Lrr44.9 are expressed during natural infection of the host and undetectable in in vitro cultures. Among all the recombinant proteins used as antigens in IgM ELISA, ArgC had the highest sensitivity and specificity, 89.8% and 95.5%, respectively, for the conclusive diagnosis of leptospirosis. The use of ArgC and RecA in combination for IgM ELISA increased the sensitivity and specificity to 95.7% and 94.9%, respectively. ArgC and RecA thus elicited specific IgM responses and were therefore effective in laboratory confirmation of Leptospira infection.

AB - Leptospirosis is a serious zoonosis that is underdiagnosed because of limited access to laboratory facilities in Southeast Asia, Central and South America, and Oceania. Timely diagnosis of locally distributed serovars of high virulence is crucial for successful care and outbreak management. Using pooled patient sera, an expression gene library of a virulent Leptospira interrogans serovar Autumnalis strain N2 isolated in South India was screened. The identified genes were characterized, and the purified recombinant proteins were used as antigens in IgM enzyme-linked immunosorbent assay (ELISA) either singly or in combination. Sera (n=118) from cases of acute leptospirosis along with sera (n=58) from healthy subjects were tested for reactivity with the identified proteins in an ELISA designed to detect specific IgM responses. We have identified nine immunoreactive proteins, ArgC, RecA, GlpF, FliD, TrmD, RplS, RnhB, Lp28.6, and Lrr44.9, which were found to be highly conserved among pathogenic leptospires. Apparently, the proteins ArgC, RecA, GlpF, FliD, TrmD, and Lrr44.9 are expressed during natural infection of the host and undetectable in in vitro cultures. Among all the recombinant proteins used as antigens in IgM ELISA, ArgC had the highest sensitivity and specificity, 89.8% and 95.5%, respectively, for the conclusive diagnosis of leptospirosis. The use of ArgC and RecA in combination for IgM ELISA increased the sensitivity and specificity to 95.7% and 94.9%, respectively. ArgC and RecA thus elicited specific IgM responses and were therefore effective in laboratory confirmation of Leptospira infection.

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