Showing 2 results for pakzad
Azar Moradkhani, Hamid Abtahi, Iraj Pakzad, Masode Karimi,
Volume 14, Issue 2 (5-2011)
Abstract
Background: Hyaluronidase A is an antigenic protein that is secreted by Streptococcus pyogenes. Nowadays, streptococcal infections are diagnosed by tracking down anti-hyaluronidase A antibodies. In this study, the attempt was made to generate recombinant hyaluronidase A in E. coli.
Materials and Methods: In this experimental study, through designing specific primers and polymerase chain reaction (PCR), hyaluronidase A gene was amplified and after purification, it was sub-cloned in plasmid expression vector pET32a. Then pET32a-hylA was transferred to E. coli BL21-DE3-plySs. Protein generation induced by IPTG. The recombinant protein was purified by Ni-NTA kit and its concentration was assayed by Bradford method. Western-Blot analysis was run for verifying the recombinant hyaluronidase A.
Results: The nucleotide sequencing of the gene amplified by PCR was the same as hyaluronidase A gene from Streptococcus pyogenes. Production of the recombinant hyaluronidase A via induction by pET32a-hylA plasmid was done through IPTG. The expressed protein was purified by affinity chromatography by Ni-NTA resin. The concentration of purified protein was 500µg/ml. analysis using a mouse anti-hyaluronidase A serum was reacted with the generated protein using Western-Blot analysis.
Conclusion: Recombinant HylA protein can be generated in E.coli and the resulting protein maintains its antigenic properties desirably.
Iraj Pakzad, Soya Bahmani, Sobhan Ghafouryan, Hassan Hosainzadegan,
Volume 14, Issue 7 (Brucellosis Supplement 2012)
Abstract
Background: Brucellosis is one of the most common zoonotic diseases in the world which imposes a great financial burden on the endemic regions. Diagnosis of the human brucellosis is mainly based on blood culture and serological tests. PCR, however, is recommended for diagnosis at greater specificities and sensitivities. This study aims to compare the diagnosis of human brucellosis by PCR method using l7/l12 and 16srRNA genes and serological tests.
Materials and Methods: In a cross-sectional study, a total of 700 blood samples were collected from patients suspected to brucellosis who had referred to the hospitals and laboratories of Ilam, Iran. The samples were selected through Rose Bengal test. Then 50 positive samples diagnosed by Rose Bengal test were assayed by Wright, Coombs Wright, and PCR using l7/l12 and 16srRNA genes and 50 negative samples diagnosed by PCR using these two genes were tested.
Results: Of the total 700 samples assayed by Rose Bengal test, 125 were positive and the rest 575 were negative. The 50 positive Rose Bengal samples in PCR were shown to be positive by both genes and 50 negative Rose Bengal samples were shown negative by both samples. 47 samples in Wright test and 49 samples in Coombs test had titration levels above 1:60.
Conclusion: PCR method has a higher sensitivity and specificity in diagnosis of human brucellosis in comparison with serological tests. Sensitivity of PCR by l7/l12 gene is similar to16srRNA and can be used for diagnosis of human brucellosis.
