Transcriptome analysis of Burkholderia pseudomallei T6SS identifies Hcp1 as a potential serodiagnostic marker

Sylvia Chieng, Rahmah Mohamed, Sheila Nathan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Burkholderia pseudomallei, the causative agent of melioidosis, is able to survive extreme environments and utilizes various virulence factors for survival and pathogenicity. To compete and survive within these different ecological niches, B.pseudomallei has evolved specialized pathways, including the Type VI secretion systems (T6SSs), that have a role in pathogenesis as well as interbacterial interactions. We examined the expression profile of B.pseudomallei T6SS six gene clusters during infection of U937 macrophage cells. T6SS-5 was robustly transcribed while the other five clusters were not significantly regulated proposing the utility of T6SS-5 as a potential biomarker of exposure to B.pseudomallei. Transcription of T6SS regulators VirAG and BprB was also not significant during infection when compared to bacteria grown in culture. Guided by these findings, three highly expressed T6SS genes, tssJ-4, hcp1 and tssE-5, were expressed as recombinant proteins and screened against melioidosis patient sera by western analysis and ELISA. Only Hcp1 was reactive by both types of analysis. The recombinant Hcp1 protein was further evaluated against a cohort of melioidosis patients (n=32) and non-melioidosis individuals (n=20) sera and the data clearly indicates a higher sensitivity (93.7%) and specificity (100%) for Hcp1 compared to bacterial lysate. The detection of anti-Hcp1 antibodies in patients' sera indicating the presence of B.pseudomallei highlights the potential of Hcp1 to be further developed as a serodiagnostic marker for melioidosis.

Original languageEnglish
Pages (from-to)47-56
Number of pages10
JournalMicrobial Pathogenesis
Volume79
DOIs
Publication statusPublished - 1 Feb 2015

Fingerprint

Melioidosis
Burkholderia pseudomallei
Gene Expression Profiling
Recombinant Proteins
Serum
U937 Cells
Virulence Factors
Multigene Family
Infection
Virulence
Anti-Idiotypic Antibodies
Biomarkers
Enzyme-Linked Immunosorbent Assay
Macrophages
Bacteria
Sensitivity and Specificity
Survival
Genes

Keywords

  • Burkholderia pseudomallei
  • Hcp1
  • Melioidosis
  • T6SS

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Transcriptome analysis of Burkholderia pseudomallei T6SS identifies Hcp1 as a potential serodiagnostic marker. / Chieng, Sylvia; Mohamed, Rahmah; Nathan, Sheila.

In: Microbial Pathogenesis, Vol. 79, 01.02.2015, p. 47-56.

Research output: Contribution to journalArticle

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abstract = "Burkholderia pseudomallei, the causative agent of melioidosis, is able to survive extreme environments and utilizes various virulence factors for survival and pathogenicity. To compete and survive within these different ecological niches, B.pseudomallei has evolved specialized pathways, including the Type VI secretion systems (T6SSs), that have a role in pathogenesis as well as interbacterial interactions. We examined the expression profile of B.pseudomallei T6SS six gene clusters during infection of U937 macrophage cells. T6SS-5 was robustly transcribed while the other five clusters were not significantly regulated proposing the utility of T6SS-5 as a potential biomarker of exposure to B.pseudomallei. Transcription of T6SS regulators VirAG and BprB was also not significant during infection when compared to bacteria grown in culture. Guided by these findings, three highly expressed T6SS genes, tssJ-4, hcp1 and tssE-5, were expressed as recombinant proteins and screened against melioidosis patient sera by western analysis and ELISA. Only Hcp1 was reactive by both types of analysis. The recombinant Hcp1 protein was further evaluated against a cohort of melioidosis patients (n=32) and non-melioidosis individuals (n=20) sera and the data clearly indicates a higher sensitivity (93.7{\%}) and specificity (100{\%}) for Hcp1 compared to bacterial lysate. The detection of anti-Hcp1 antibodies in patients' sera indicating the presence of B.pseudomallei highlights the potential of Hcp1 to be further developed as a serodiagnostic marker for melioidosis.",
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