Extraction and Purification of Haemophilus influenzae Type b Lipooligosac‌charide by Modified Phenol Method

Authors

  • Ahmad Reza Bahremand Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Ali Nour Neamatollahi Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Amin Arsang Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran, Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Farzam Vaziri Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Mehdi Nejati Department of Bacterial vaccine, Pasteur Institute of Iran, Tehran, Iran
  • Morteza Masoumi Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Seyed Davar Siadat Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran, Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
  • Shamsi Yari Department of Mycobacteriology & Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran
Abstract:

  Introduction : Haemophilus influenzae type b (Hib) is a Gram negative bacterium and one of the causative agents of acute bacterial meningitis, especially in infants and children less than 5 years old. Lipooligosaccharide (LOS), one of the virulence factors which plays an important role in pathogenesis of Hib, has multiple applications in diagnosis and conjugate vaccines. In this study, LOS extracted from two Hib standard strains (ATCC 39930 and ATCC 10214) were compared. Material and methods: LOS was extracted by a modified hot phenol method from the aqueous and phenol phase and its concentration and purity assayed . Protein c ontaminations of the samples were determined spectrophotometrically and their endotoxin contents were assayed by the Limulus amebocyte lysate (LAL). Result and Discussion: The yield of the extracted LOS from strain ATCC10214 was about 475 μg/ml and the protein contaminations of the samples were approximately 0.07 mg/ml, whereas strain ATCC39930 yielded 520 μg/ml of LOS with protein contamination of 0.08 mg/ml. The results showed that the production of LOS by both strains was similar and the variation observed was not statistically significant (p < 0.001). Vac Res , 2014, 1 (1): 29-31

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Journal title

volume 1  issue 1

pages  28- 30

publication date 2014-08

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