FIELD: medicine.
SUBSTANCE: method involves applying polymerase chain reaction diagnostic methods for detecting perinatal infection pathogenic agents in afterbirth. Preparation for detecting DNA- and RNA-containing pathogenic agents, is obtained as follows. 1.0x1.0 cm large placenta samples are taken from fetal membrane rupture places and in intervillous lacuna in the afterbirth center. The samples are placed into reservoir filled with sterile physiologic saline to be kept in storage not longer than 4 h at+4°C temperature. Then the samples are grinded and alkaline extraction is carried out with 200 mcl adding 200 mcl of solution containing 1.5 M NaCl, 0.3 M NaOH, 0.1% Sacrosyl solution and 0.5% sodium dodecylsulfate. The sample is left for 16-18 h at temperature of +4°C. Then the suspension is shaken, centrifuged at 5000 rpm during 5 min. The so produced suspension is transferred into test-tube having volume of 1.5 ml and extraction with chloroform and phenol and denaturation with ethanol is carried out.
EFFECT: high accuracy of measurements.
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Authors
Dates
2007-01-27—Published
2005-09-26—Filed