Primer antitüberküloz ilaçlara dirençli mycobacterium tuberculosis kompleks izolatlarının moleküler yöntemlerle genotiplendirilmesi
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Tarih
2022
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Yayıncı
Dicle Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Fenotipik olarak primer anti-tüberküloz ilaçlara dirençli olduğu saptanan Mycobacterium tuberculosis kompleks (MTBK) izolatlarının spoligotipleme, MIRU-VNTR (Mycobacterial Interspersed Repetitive Units –Variable Number of Tandem Repeats) yöntemleriyle genotiplendirilmesi ve dirence neden olan mutasyonların tespit edilmesi amaçlandı. Gereç ve Yöntem: Çalışmaya Ocak 2014-Ağustos 2019 tarihleri arasında Dicle Üniversitesi Hastanesi Laboratuvarı Mikobakteriyoloji biriminde klinik materyallerden izole edilen ve MGIT 960 sistemi ile primer antitüberküloz ilaçlara dirençli olduğu saptanan MTBK izolatları dahil edildi. Toplam 69 izolatın 32'si (%46.4) çoklu ilaç dirençli (ÇİD), 34'ü (%49.3) izoniazid (INH) ile birlikte primer anti-tüberküloz ajanlardan bir veya birkaçına, üçü (%4.3) ise rifampine (RIF) dirençliydi. İzolatlar spoligotipleme ve 15 lokus MIRU-VNTR yöntemleri ile genotipik çeşitlilik ve klonal ilişkiler yönünden değerlendirildi, GenoType MTBDR plus (Hain Lifescience, Almanya) kiti ile dirençle ilişkili gen mutasyonları saptandı. Bulgular: Spoligotipleme sonucu en çok üyeye sahip aileler T1 (%55.4) ile TUR (%13.8) idi. İzolatların yaklaşık üçte biri (%36.92) T1 ailesinin SIT53'ünde, %9.23'ü SIT284, %12.31'i SIT41'de ve %0.77'si herhangi bir aileye dahil olmayan spoligotip SIT2560'ta kümelendi. Yedi izolat SIT2560, birer izolat SIT4, SIT608 ve bir izolat da orphan suş olarak tanımlandı. RIF dirençli izolatların 19'unda (%57.6) saptanan, rpoB geninin 531. Gen bölgesindeki TCG/TTG mutasyonu dirençten sorumlu en yaygın mutasyondu. INH dirençli izolatların 14'ünde (%46.7) katG315 gen bölgesinde AGC/ACC mutasyonu, 8'inde (%26.7) ise inhA gen bölgesinin -15 pozisyonunda C/T mutasyonları saptandı. Sonuç: İzolatlarımızın kümelendiği majör familyaların %55.4 ile T1 ve %13.8 ile TUR olduğu, saptanan kümelerin genellikle farklı illerden gelen hastalara ait karışık izolatlardan oluştuğu ve aralarında klonal ilişki bulunmadığı, dirençli izolatların literatürde belirtilen yaygın mutasyonlar gösterdikleri saptandı.
Aim: Genotype determination of Mycobacterium tuberculosis complex (MTBC) isolates, which were phenotypically resistant to primary anti-tuberculosis drugs, by spoligotyping and Mycobacterial interspersed repetetive units, (MIRU) - variable number of tandem repeat (VNTR) typing methods was aimed. Materials and Methods: MTBC strains isolated from clinical materials in Dicle University Hospital Laboratory Mycobacteriology unit and found to be resistant to primary antituberculosis drugs with the MGIT 960 system between January 2014 and August 2019 were included. Of the 69 isolates, 32 (46.4%) were multi-drug resistant (MDR), 34 (49.3%) were resistant to one or more of the primary anti-tuberculosis agents in addition to isoniazid (INH), and three (4.3%) were resistant to rifampin (RIF). Isolates were evaluated in terms of genotypic diversity and clonal associations by spoligotyping and 15 loci MIRU-VNTR methods. Resistance genes were determined by GenoType MTBDR plus (Hain Lifescience, Germany). Results: Families with the highest number of members by spoligotyping were T1 (55.4%) and TUR (13.8%). Approximately one-third (36.92%) of the isolates clustered in SIT53 of T1 family, 9.23% in SIT284, 12.31% in SIT41, and 0.77% in the spoligotype SIT2560, which was not included in any family. Seven isolates were identified as SIT2560, one isolate SIT4, SIT608 and orphan strain. TCG/TTG mutation in the 531st gene region of the rpoB gene, found in 19 (57.6%) of the RIF resistant isolates, was the most detected mutation responsible for resistance. AGC/ACC mutations were found in the katG315 gene region in 14 (46.7%) of the INH resistant isolates, and C/T mutations were detected in the -15 position of the inhA gene region in 8 (26.7%) of the INH resistant isolates. Conclusion: The major families in which our isolates clustered were T1 with 55.4% and TUR with 13.8%, the clusters determined were generally mixed isolates from patients from different provinces and there was no clonal relationship between them, and resistant isolates showed common mutations stated in the literature.
Aim: Genotype determination of Mycobacterium tuberculosis complex (MTBC) isolates, which were phenotypically resistant to primary anti-tuberculosis drugs, by spoligotyping and Mycobacterial interspersed repetetive units, (MIRU) - variable number of tandem repeat (VNTR) typing methods was aimed. Materials and Methods: MTBC strains isolated from clinical materials in Dicle University Hospital Laboratory Mycobacteriology unit and found to be resistant to primary antituberculosis drugs with the MGIT 960 system between January 2014 and August 2019 were included. Of the 69 isolates, 32 (46.4%) were multi-drug resistant (MDR), 34 (49.3%) were resistant to one or more of the primary anti-tuberculosis agents in addition to isoniazid (INH), and three (4.3%) were resistant to rifampin (RIF). Isolates were evaluated in terms of genotypic diversity and clonal associations by spoligotyping and 15 loci MIRU-VNTR methods. Resistance genes were determined by GenoType MTBDR plus (Hain Lifescience, Germany). Results: Families with the highest number of members by spoligotyping were T1 (55.4%) and TUR (13.8%). Approximately one-third (36.92%) of the isolates clustered in SIT53 of T1 family, 9.23% in SIT284, 12.31% in SIT41, and 0.77% in the spoligotype SIT2560, which was not included in any family. Seven isolates were identified as SIT2560, one isolate SIT4, SIT608 and orphan strain. TCG/TTG mutation in the 531st gene region of the rpoB gene, found in 19 (57.6%) of the RIF resistant isolates, was the most detected mutation responsible for resistance. AGC/ACC mutations were found in the katG315 gene region in 14 (46.7%) of the INH resistant isolates, and C/T mutations were detected in the -15 position of the inhA gene region in 8 (26.7%) of the INH resistant isolates. Conclusion: The major families in which our isolates clustered were T1 with 55.4% and TUR with 13.8%, the clusters determined were generally mixed isolates from patients from different provinces and there was no clonal relationship between them, and resistant isolates showed common mutations stated in the literature.
Açıklama
Anahtar Kelimeler
Mycobacterium tuberculosis, İlaç direnci, Genotipleme teknikleri, Mycobacterium tuberculosis, Drug resistance, Genotyping techniques
Kaynak
WoS Q Değeri
Scopus Q Değeri
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Künye
Şahin, İ. H. (2022). Primer antitüberküloz ilaçlara dirençli mycobacterium tuberculosis kompleks izolatlarının moleküler yöntemlerle genotiplendirilmesi. Yayımlanmamış doktora tezi, Dicle Üniversitesi, Diyarbakır.
