Open AccessEFFICIENCY OF APPLICATION OF ISOTHERMAL AMPLIFICATION AT INSPECTION OF PATIENTS WITH WHOOPING COUGH
Author(s) -
А. С. Пименова,
O. Yu. Borisova,
М. С. Петрова,
I. S. Voronina,
А. Б. Борисова,
О. В. Шамшева,
S. S. Afanasiev,
Е. В. Власов,
В. А. Алешкин
Publication year - 2018
Publication title -
infekciâ i immunitet
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.137
H-Index - 5
eISSN - 2313-7398
pISSN - 2220-7619
DOI - 10.15789/2220-7619-2018-3-361-368
Subject(s) - whooping cough , loop mediated isothermal amplification , medicine , dna extraction , bordetella pertussis , dna , pediatrics , polymerase chain reaction , pathology , biology , bacteria , genetics , gene , vaccination
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%.