EFFICIENCY OF APPLICATION OF ISOTHERMAL AMPLIFICATION AT INSPECTION OF PATIENTS WITH WHOOPING COUGH

Abstract

Purpose: efficiency isothermal amplification (LAMP) at inspection of patients with whooping cough in clinical conditions. Materials and methods. Examination of 262 patients aged from 0 months up to 30 years hospitalized in Infectious diseases clinical hospital No. 1 of the Moscow Department of Healthcare is conducted. Clinical specimens (pharyngeal swabs) were collected according to MR 3.1.2.0072-13. Extraction DNA of B. pertussiswas carried out by means of the АmplyPrime® DNA-sorb-AM. Identification of specific fragments of a genome of B. pertussiswas performed by PCR-real time by means of the АmplySens ® Bordetella Multi-FL set (a comparison method) and by the LAMP with a phoresis and intercalating dye. Results.When using of the optimized method LAMP the DNA of B. pertussisis found at 252 (96.2%) patients. The method was effective at any forms of whooping cough —DNA of B. pertussisis found in all patients with a severe form, in 95.8% of cases — in patients with medium-weight and in 95.3% of cases — in patients with an easy form. The DNA of B. pertussisis found in clinical specimens received from patients on different terms from the beginning of a disease — from 92.3% on the 1st week up to 96% of cases — on the 5 th and more weeks of a disease. The DNA of B. pertussisis found in high percent of cases (96.7–95.9%) and did not depend on acceptance of antibacterial therapy. Children till 1 year are the main age group which is subject to hospitalization at suspicion of whooping cough and in which the highest risk of development of complications and severe forms of a clinical disease. At inspection of 169 children from 0 to 12 months by means of the optimized method LAMP, DNA of B. pertussisit is found in 98.6% of cases in children of patients with whooping cough aged from 0–3 months, in 98.4% of cases — children of 4–7 months and in 94.6% of cases — at children have 8–12 months. The efficiency of detection of DNA of B. pertussisat patients with whooping cough of children aged till 1 year by means of the optimized LAMP method was 97.6%.