Truncation of PstS1 antigen of Mycobacterium tuberculosis improves diagnostic efficiency

Fusion of selected regions of mycobacterial antigens for enhancing sensitivity in serodiagnosis of tuberculosis

“…Similarly, 126 bp fragment of Rv3803c (which encodes tn2FbpC1 fragment i.e., 97-111 amino acid residues of FbpC1) was amplified using the forward primer F2 and the reverse primer R1. PCR was also done to amplify 792 bp fragment of Rv0934 (which encodes tnPstS1 i.e., 97-360 amino acid residues of PstS1) as described previously (Khurshid et al, 2013). This fragment was ligated into EcoRI and HindIII sites of pET28a(+).…”

“…Full length Rv0251c, encoding HSP, was amplified using the primers F5 and R5. Full length Rv0934, encoding PstS1 was amplified as described previously (Khurshid et al, 2013). All the full length genes were initially cloned in pTZ57R/T and then subcloned in the vector pET28a(+).…”

“…The proteins were further purified by anion exchange FPLC using HiTrap Q FF 5 ml column (GE Healthcare). PstS1 and tnPstS1 were purified using single step purification and refolding method as described previously (Khurshid et al, 2013).…”

“…Institutional animal experimentation guidelines and EU Directive 2010/63/EU for animal experiments were followed. All the procedures were the same as reported previously (Khurshid et al, 2013).…”

“…We have shown previously that the truncation of PstS1, by removing 96 and 14 amino acid residues from the N-and C-terminals respectively, improves its diagnostic efficiency (Khurshid et al, 2013). This study describes the sensitivity and specificity of the novel fusion proteins constructed from the truncated versions of PstS1, FbpC1 and HSP for diagnosis of active TB patients.…”