ГЕНЕТИЧЕСКИЕ ОСОБЕННОСТИ ВАКЦИННОГО СУБШТАММА BCG RUSSIA В КОНТРОЛЕ ПРОИЗВОДСТВА И ПОСТВАКЦИНАЛЬНЫХ ОСЛОЖНЕНИЙ, "Вестник Центрального научно-исследовательского института туберкулеза"

2017 ◽  
pp. 22-31
Author(s):  
О.Л. Воронина ◽  
М.С. Кунда ◽  
Е.И. Аксенова ◽  
Н.Н. Рыжова ◽  
А.Н. Семенов ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Expert Committee ◽  
Accession Number ◽  
Biological Standardization

Введение БЦЖ (Бацилла-Кальмета-Герена) вакцина - единственная разрешенная к применению, позволяющая противостоять Mycobacterium tuberculosis. Разработка международных требований к производству и контролю БЦЖ вакцины находится под контролем экспертного комитета ВОЗ по биологической стандартизации (Expert Committee on Biological Standardization, ECBS). В 2009 г. ECBS учредил референсы ВОЗ для БЦЖ вакцины на основе трех субштаммов: Danish1331, Tokyo 172-1 и Russian BCG-I, а также требования к контролю, включающие генетическую характеристику произведенных и посевных серий вакцинного субштамма. Согласно требованиям ВОЗ и надлежащей производственной практики (GMP) было выполнено секвенирование трех посевных серий BCG Russia на платформе 454 Roche в нашей лаборатории. БЦЖиты (лимфадениты, остеомиелиты - осложнения при вакцинации БЦЖ) составляют 1% у здоровых реципиентов, однако занимают третье место среди поствакцинальных осложнений в Российской Федерации. Цель наших исследований заключалась в выявлении генетических особенностей субштамма BCG Russia для разработки экспресс-типирования при осложнениях. Методы Полногеномное секвенирование на платформе 454 Roche. MIRU-VNTRplus database для анализа отличий в профилях BCG субштаммов. Набор биоинформационных ресурсов для аннотирования и исследования геномов. Результаты Геном BCG Russia 368 был секвенирован и депонирован в GenBank (Accession Number NZ_CP009243.1). Сравнение геномов последней генерации субштамма и двух предшествующих продемонстрировало стабильность субштамма BCG Russia. Отличие от ближайшего раннего субштамма BCG Tokyo 172 включало 2 RD (region of difference): инсерцию 22 bp и делецию 1602 bp, десять ins/del (1-9 bp) и 52 SNP. Профиль выявленных профагов в геномах M. bovis и M. bovis BCG отражал эволюцию вакцинных субштаммов. Геномы M. bovis содержали профаги 7.5 и 20.3 kb, ранние BCG субштаммы имели профаги 7.5 and 11.2 kb. Большинство поздних субштаммов утратило профаг 11.2 kb. Исключение составили BCG Montreal and BCG Tice, в геномах которых выявлено 6 и 15 профагов, соответственно. Анализ 27 MIRU-VNTR локусов в геноме BCG Russia 368, их сравнение с профилями локусов штаммов M. bovis и BCG субштаммов в MIRU-VNTRplus database позволили обосновать специфичность семи локусов для субштамма BCG Russia. Мы выбрали Mtub21 и Mtub04 - локусы без повторов в геноме BCG Russia, Mtub-30, QUB-26, QUB-323222 -наиболее вариабельные среди M. bovis и M. bovis BCG, и VNTR-3820 и VNTR-4120 как гипервариабельные локусы. Фрагмент анализ этих локусов позволит провести экспресс тестирование при БЦЖитах. Заключение Полногеномное секвенирование - успешный подход в сравнении близкородственных субштаммов и в выявлении специфических локусов для диагностической панели.

scholarly journals Analisis Mutasi pada Kodon 531 Pada Gen Rpob Mycobacterium tuberculosis Penyebab Resistensi Rifampisin

2017 ◽  
Vol 15 (2) ◽  
pp. 140
Author(s):  
Yatnita Parama Cita ◽  
Dwi Hilda Putri
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Bacterial Resistance ◽  
Rpob Gene ◽  
Resistance Mechanisms ◽  
Primer Design ◽  
Reference Sequence ◽  
Multi Drug Resistance ◽  
Accession Number ◽  
Chemotherapy Agent ◽  
Primer Design Program

Tuberculosis (TB) is a serious disesase in the world. According to the WHO, it is estimated more than 3 million people die every year as a result of this infectious disease. One factor that causes diffi culty handling TB chemoteraphy is not effective against the bacteria Mycobacterium tuberculosis that causes TB . Effectiveness of treatment is often hampered by the emergence of bacterial resistance against M. Tuberculosis chemotherapy agents are given. From some research found that bacterial resistance may occur in more one type of chemotherapy agent also known as multi-drug resistance (MDR). Mycobacterium tuberculosis develop resistance mechanisms that are different from other bacteria in general. In prokaryotes, resistance is generally due to the transfer of genetic, either through plasmids,transposons and other. Reference sequence beta sub unit of RNAP protein M. Tuberculosis with accession number NP_215181.1 and M. tucerculosis rpoB gene with accession number NC_000962.3 used to obtain preliminary information from the data base www.ncbi.nlm.gov and www.uniprot.org . Mutation done according to several studies literature. Analysis of the composition, profi le, location and structure of protein using www.expasy.org, TMHMM and http://bioinf.cs.ucl.ac.uk/psipred. The primer design is done with Primer Design Program. Based on the analysis of mutation in the beta subunit of RNAP protein M. Tuberculosis, codon 531 (Ser ->Leu), it is known that mutations cause changes in some properties and structure of proteins. Possible changes affecting the nature of bacterial resistance to antibiotics rifampicin. However, further analysis needs to be done with the analysis of the docking technique.

scholarly journals International collaborative study to evaluate a recombinant L ferritin preparation as an International Standard

1997 ◽  
Vol 43 (9) ◽  
pp. 1582-1587 ◽  
Author(s):  
Susan J Thorpe ◽  
Dawn Walker ◽  
Paolo Arosio ◽  
Alan Heath ◽  
James D Cook ◽  
...  
Keyword(s):  
Collaborative Study ◽  
Expert Committee ◽  
Immunological Reactivity ◽  
International Standard ◽  
Accelerated Degradation ◽  
Wide Range ◽  
Biological Standardization ◽  
International Collaborative ◽  
Degradation Studies ◽  
International Collaborative Study

Abstract A recombinant L ferritin preparation, lyophilized in ampoules and designated 94/572, was evaluated by 18 laboratories in 9 countries for its suitability as an International Standard (IS). The preparation was assayed in a wide range of in-house and commercial immunoassays against the 2nd IS for ferritin (of spleen origin; 80/578). The immunological reactivity of the recombinant material was similar to that of the 2nd IS for ferritin in the majority of assays and demonstrated adequate stability in accelerated degradation studies. On the basis of the results presented here, the WHO Expert Committee on Biological Standardization established 94/572 as the 3rd IS for ferritin, recombinant.

THE INTERNATIONAL REFERENCE PREPARATION OF HUMAN PITUITARY LUTEINIZING HORMONE, FOR IMMUNOASSAY

1978 ◽  
Vol 88 (2) ◽  
pp. 250-259 ◽  
Author(s):  
P. L. Storring ◽  
D. R. Bangham ◽  
P. Mary Cotes ◽  
Rose E. Gaines ◽  
S. L. Jeffcoate
Keyword(s):  
Luteinizing Hormone ◽  
Reproductive Organ ◽  
Expert Committee ◽  
Confidence Limits ◽  
Binding Assays ◽  
Human Pituitary ◽  
International Reference ◽  
Biological Standardization ◽  
Reference Preparation

ABSTRACT The preparation and nature of the International Reference Preparation of Human Pituitary Luteinizing Hormone for Immunoassay are described. A collaborative assay of this material (coded 68/40) in terms of the International Reference Preparation of Human Pituitary Gonadotrophins (FSH and LH (ICSH)), for Bioassay (coded 69/104) was carried out by twelve laboratories in seven different countries, using different bioassay and immunoassay methods. The weighted combined potency estimate (with 95 % confidence limits) was 52.1 (46.3–58.7) IU/ampoule with male accessory reproductive organ weight gain assays; 80.2 (73.2–87.9) IU/ampoule with ovarian ascorbate depletion assays; 127 (124–129) IU/ampoule with in vitro Leydig cell testosterone production assays; and 124 (121–126) IU/ampoule with testis receptor binding assays. Immunoassay estimates in terms of the same standard were heterogeneous and gave an unweighted mean potency estimate of 33.2 IU with 95% confidence limits of 14.8– 74.4 IU/ampoule. Estimates from different methods gave significantly different results, and the reasons for this are discussed in terms of the differences between the materials being compared and the methods used in the comparison. These data illustrate the conceptual difficulties involved in comparing hetero geneous reference preparations, especially by both bioassay and immunoassay, and some of the causes of inevitable discontinuity of assay results, as described in the 26th Report of the WHO Expert Committee on Biological Standardization. On the basis of these results, and in the interest of maintaining continuity of its unitage, the International Reference Preparation has been allocated a potency of 77 IU/ampoule.

scholarly journals International collaborative study to evaluate and calibrate two recombinant L chain Ferritin preparations for use as a WHO International Standard

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Bernard Fox ◽  
Graham Roberts ◽  
Eleanor Atkinson ◽  
Peter Rigsby ◽  
Christina Ball
Keyword(s):  
Geometric Mean ◽  
Collaborative Study ◽  
Expert Committee ◽  
International Standard ◽  
Long Term Storage ◽  
Automated Assay ◽  
Accelerated Stability ◽  
Biological Standardization ◽  
International Collaborative ◽  
International Collaborative Study

Abstract Objectives To evaluate and calibrate two candidate preparations for the 4th International Standard for Ferritin (Human, Recombinant) (codes: 19/118 and 19/162) against the 3rd International Standard for Ferritin (Human, Recombinant) (code: 94/572), and three serum commutability samples in an international collaborative study involving 12 laboratories in nine countries. Methods Eleven of the 12 participating laboratories performed Ferritin quantitation using automated assay platforms and one laboratory used a manual ELISA kit. Results There was better overall agreement between all laboratories and between assay methods for the potency of preparation 19/118 than for preparation 19/162. The overall geometric mean potency (from all methods) of the candidate 4th International Standard, 19/118, was 10.5 µg/ampoule, with inter-laboratory variability, expressed as % geometric coefficient of variation (GCV), of 4.7%. Accelerated stability studies have predicted both 19/118 and 19/162 to be very stable for long term storage at −20 °C. Conclusions The candidate 4th International Standard for Ferritin (Human, Recombinant) (19/118) has been shown to be immunologically similar to the 3rd International Standard for Ferritin (Human, Recombinant) (94/572). It was recommended to and accepted by the WHO Expert Committee on Biological Standardization that 19/118 be established as the 4th International Standard for Ferritin (Human, Recombinant) with an assigned potency of 10.5 µg/ampoule and expanded uncertainty limits 10.2–10.8 µg/ampoule (95% confidence; k=2.23).

Calibration of Reference Thromboplastins and Standardisation of the Prothrombin Time Ratio

1983 ◽  
Vol 49 (03) ◽  
pp. 238-244 ◽  
Author(s):  
T B L Kirkwood
Keyword(s):  
Prothrombin Time ◽  
Oral Anticoagulants ◽  
Collaborative Study ◽  
World Health ◽  
Expert Committee ◽  
Calibration Model ◽  
Biological Standardization ◽  
Reference Preparation ◽  
Health Organization ◽  
Practical Implications

SummaryThromboplastins vary in their sensitivity to the haemostatic defect induced by oral anticoagulants. To provide a means of standardising prothrombin time tests, the World Health Organization adopted in 1977 a scheme for calibrating thromboplastins in terms of an International Reference Preparation. Unfortunately, the model on which this scheme was based does not always hold. A revised calibration model has therefore been developed and this has been tested in a recent collaborative study. The revised model, which retains fundamentally the same principle for standardising prothrombin time tests, has proved suitable for calibrating thromboplastins of different species and types and, moreover, has certain statistical advantages over its predecessor. In September 1982, the WHO Expert Committee on Biological Standardization adopted the revised model. This paper explains the nature and rationale of this change and considers its practical implications.

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