International Medical Journal, Vol. 21., Iss. 3, 2015, P. 74−76.
QUALITATIVE AND QUANTITATIVE DETERMINING OF PEPTIDOGLYCAN IN PATIENTS WITH PURULENT BACTERIAL MENINGITIS BY PLASMA OF SILKWORM LARVAE
Kharkiv Medical Academy of Postgraduate Education, Ukraine
The aim of this work was to develop a biological method for qualitative and quantitative determination of peptidoglycan (PG) in patients with purulent bacterial meningitis (PBM) using hemolymph of the silkworm (SLP). SLP−reaction was used due to the following reasons. Hemolymph of the silkworm (Воmbix mori ?.) contains natural enzyme prophenol−oxidase that acts as a catalyst for the reaction of interaction of PG with exogenous substrate 3,4−digarettes with formation of melanin. The activity of the enzyme, and therefore, the color intensity is proportional to the concentration of GHGs. Proposed by Japanese scientists in 2003 SLP−test was piloted on a small number of patients; the study did not involve the patients with meningococcal meningitis, the causative agent of which is the main etiological agent in the structure of PBM. Our study involved 38 patients with acute meningitis of different etiology and 10 controls. Live clonally «population» of elite В. mori ?, was obtained from V.N. Karazin University laboratory of embryonic and stem cells. SLP−test of CSF (cerebrospinal fluid) of patients with confirmed diagnosis of bacterial meningitis demonstrated dark coloration of the investigated mixtures, and in patients with viral meningitis and controls coloring of mixtures did not occur. To determine the concentration of PGs in the CSF of the patient according to the degree of coloring, preliminary constructed calibration curve was used. In the acute phase and the recovery period as well as in the dynamics of the disease, the concentration of the studied endotoxin in the CSF of patients with pneumococcal meningitis was statistically higher than in patients with meningococcal meningitis. The results confirm a greater availability of PG in patients with pneumococcal meningitis than in meningococcal meningitis, where the content of PG is insignificant. Visual distinguishing by the intensity of staining of gram−positive pneumococcal from gram−negative meningococcal meningitis was not possible. Qualitative and quantitative SLP−test can be used for early diagnosis and differential diagnosis of acute meningitis, and also for monitoring the effectiveness of antibiotic therapy in patients PBM. In favour of the introduction into clinical practice of innovative ideas is the presence in Ukraine of the Institute of Sericulture of the Ukrainian Academy of Agrarian Sciences, cultivating the silkworm.
Key words: peptidoglycan, plasma of silkworm larvae, cerebrospinal fluid, acute bacterial meningitis.