The purpose of study is to develop techniques of laboratory diagnostic of leprosy on the basis of polymerase chain reaction. The techniques were worked through such as extraction of DNA from various clinical material (bioptates and scarifcate of skin, scrapes from mucous surface of nose and trophic ulcers, blood serum). The systems of oligonucleotides were constructed to 16S pRNA and probe for identifcation of Mycobacterium leprae in real-time format for amplifer DT-96 ("The DNA-Technology", Russia). The optimal regimen of amplifcation was worked out. To control polymerase chain reaction processing a system of internal control was introduced. The evaluation of specifcity of test-system was implemented using 17 strains of various types of mycobacteria and clinical samples from 32 patients with leprosy and 15 healthy individuals being in family contact with patients with leprosy. The 100% sensitivity was established concerning detection of Mycobacterium leprae in bioptates of skin using polymerase chain reaction technique as compared with standard bacterioscopic technique (88.9%). In all other clinical samples, a higher sensitivity of developed test-system on the basis of polymerase chain reaction was detected too. The proposed test-system has higher sensitivity and specifcity that permits to resolve an issue of fast identifcation of Mycobacterium leprae. It can be applied in epidemiological studies at investigation of prevalence of agent of disease.
BT - Klinicheskaia laboratornaia diagnostika C1 -http://www.ncbi.nlm.nih.gov/pubmed/30550093?dopt=Abstract
DO - 10.18821/0869-2084-2018-63-1-55-59 IS - 1 J2 - Klin. Lab. Diagn. LA - rus N2 -The purpose of study is to develop techniques of laboratory diagnostic of leprosy on the basis of polymerase chain reaction. The techniques were worked through such as extraction of DNA from various clinical material (bioptates and scarifcate of skin, scrapes from mucous surface of nose and trophic ulcers, blood serum). The systems of oligonucleotides were constructed to 16S pRNA and probe for identifcation of Mycobacterium leprae in real-time format for amplifer DT-96 ("The DNA-Technology", Russia). The optimal regimen of amplifcation was worked out. To control polymerase chain reaction processing a system of internal control was introduced. The evaluation of specifcity of test-system was implemented using 17 strains of various types of mycobacteria and clinical samples from 32 patients with leprosy and 15 healthy individuals being in family contact with patients with leprosy. The 100% sensitivity was established concerning detection of Mycobacterium leprae in bioptates of skin using polymerase chain reaction technique as compared with standard bacterioscopic technique (88.9%). In all other clinical samples, a higher sensitivity of developed test-system on the basis of polymerase chain reaction was detected too. The proposed test-system has higher sensitivity and specifcity that permits to resolve an issue of fast identifcation of Mycobacterium leprae. It can be applied in epidemiological studies at investigation of prevalence of agent of disease.
PY - 2018 SP - 55 EP - 59 T2 - Klinicheskaia laboratornaia diagnostika TI - [The development of laboratory diagnostic of leprosy using polymerase chain reaction]. VL - 63 SN - 0869-2084 ER -