Lack of serological specificity of recombinant heat shock protein of Leishmania donovani

S. K. Arora, Peter Melby, S. Sehgal

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In order to identify a specific recombinant antigen of Leishmania donovani with potential use for diagnosis, a cDNA library was constructed in lambda ZAP II expression vector. On screening the cDNA library using pooled sera from Indian patients with kala azar, 20 antibody reactive clones were identified. These were subcloned into pBluescript phagemid by an in vivo excision procedure. The molecular weights of the expressed recombinant proteins varied from 15 to 70 kDa and the cDNA insert sizes varied from 0.5 kb to the largest size of approximately 2.0 kb which was designated as the E2b clone. The nucleotide sequencing revealed that 50% of the clones had sequence homology to the heat shock protein gene of L. donovani. The serological studies conducted with the kala azar positive sera and sera from healthy laboratory workers using the recombinant protein from the E2b clone and having sequence homology to Ldhsp 70, indicated that although all the kala azar sera were positive, 12 of 20 healthy individuals also showed antibodies against the recombinant hsp70, indicating that this antigen is not suitable for serological diagnosis of kala azar.

Original languageEnglish (US)
Pages (from-to)446-451
Number of pages6
JournalImmunology and Cell Biology
Volume73
Issue number5
StatePublished - 1995
Externally publishedYes

Fingerprint

Leishmania donovani
Visceral Leishmaniasis
Heat-Shock Proteins
Gene Library
Recombinant Proteins
Clone Cells
Antigens
Antibodies
Sequence Homology
Serum
Screening
Nucleotides
Complementary DNA
Genes
Molecular weight
Molecular Weight

Keywords

  • cDNA inserts
  • cDNA library
  • Heat shock protein
  • Lambda ZAP-II vector
  • Leishmania donovani
  • Phagemid
  • Plasmids
  • Recombinant fusion proteins
  • Western blot

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Immunology

Cite this

Lack of serological specificity of recombinant heat shock protein of Leishmania donovani. / Arora, S. K.; Melby, Peter; Sehgal, S.

In: Immunology and Cell Biology, Vol. 73, No. 5, 1995, p. 446-451.

Research output: Contribution to journalArticle

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