Investigation of phenotypic and genotypic characteristics of carbapenem resistant Klebsiella pneumoniae strains isolated from nosocomial infections

Year 2020, , 91 - 99, 21.01.2020

Sema Alaçam , İrmak Baran

https://doi.org/10.31362/patd.621807

Abstract

Purpose: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an
important nosocomial pathogen. The most common mechanism that causes carbapenem
resistance in these bacteria is carbapenemase production. We aimed to study the
molecular epidemiology of these K.
pneumoniae isolates and the characteristics of carbapenem resistance.

Materials and Methods: The
diagnosis and antibiotic susceptibility tests of 62 CRKP isolates were
performed with Vitek-2 automated system and E test. Carbapenemase genes were
determined by multiplex PCR. The genetic relationships were detected by rep-PCR.

Results:
Eight
clusters containing 25 strains with 95% or more similarity were identified by
Rep-PCR method. On the other hand, 59.7% (37/62) of these strains were
identified as "different" with similarities of less than 95% and more
than two different bands. The blaOXA-48 gene was detected in 57 of the 62
isolates (91.9%) and the modified Hodge test was positive in all of the blaOXA-48
gene positive isolates. The resistance rates of carbapenems (imipenem,
meropenem and ertapenem) were examined by Vitek-2 system and E-test. The lowest
resistance rates were observed for meropenem (Vitek-2 35%, E-test 16%).  Resistance rates in these isolates were 100%
for ampicillin and amoxicillin-clavulanic acid, 90% for ceftriaxone.

Conclusion: We found that OXA-48 oxacillinase production was
responsible for carbapenemase resistance in CRKP. The rep-PCR results suggested
that the genetic similarities between CRKP isolates were not high. We suggest
that this was due to the characteristic of blaOXA-48 gene which is horizontal
trasnferred rather. than clonal spread among bacteria. 

Keywords

OXA-48, carbapenemase, rep-PCR, multiplex PCR

Hastane enfeksiyonlarından izole edilen karbapenem dirençli Klebsiella pneumonia suşlarının fenotipik ve genotipik özelliklerinin incelenmesi

Abstract

Amaç: Karbapenemlere
dirençli Klebsiella pneumoniae (KDKP)
önemli bir nosokomiyal patojendir. Bu bakterilerde karbapenem direncine neden
olan en yaygın mekanizma karbapenemaz üretimidir. Bu çalışmada KDKP
izolatlarının moleküler epidemiyolojisini ve karbapenem direncinin
özelliklerini incelemeyi amaçladık.

Gereç
ve Yöntem: Toplam 62 KDKP izolatının tanımlanması ve
antibiyotik duyarlılık testleri Vitek-2 otomatik sistem ve E testi ile yapıldı.
Karbapenemaz genleri multipleks PCR ile belirlendi. Genetik benzerlikleri
rep-PCR ile araştırıldı.

Bulgular:
Rep-PCR
yöntemi ile, %95 veya daha fazla benzerliğe sahip 25 suş içeren sekiz küme
tanımlandı. İzolatların %59.7'si (37/62), benzerlikleri %95'ten az ve ikiden
fazla farklı bant içeren, “farklı” olarak belirlendi. BlaOXA-48 geni,
izolatların %91.9’unda (57/62) tespit edildi. Modifiye Hodge testi, blaOXA-48
geni pozitif izolatlarının tamamında pozitifti. Karbapenemlerin (imipenem,
meropenem ve ertapenem) direnç oranları Vitek-2 sistemi ve E-testi ile
incelendi. En düşük direnç oranları meropenem için belirlendi (Vitek-2 %35,
E-test %16). Bu izolatlarda direnç oranları ampisilin ve amoksisilin-klavulanik
asit için %100, seftriakson için %90 bulundu.

Sonuç:
KDKP
izolatlarında OXA-48 oksasilinaz üretiminin karbapenemaz direncinden sorumlu
olduğunu belirledik. Rep-PCR sonuçları, KDKP izolatları arasındaki genetik
benzerliklerin yüksek olmadığını göstermiştir. Bu verilere dayanarak, K. pneumoniae suşlarında blaOXA-48
geninin, klonal yayılımdan ziyade bakteriler arasında horizontal geçiş
özelliğinden kaynaklandığını ileri sürmekteyiz.

Keywords

OXA-48, karbapenemaz, rep-PCR, multipleks PCR

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