{"id":1504,"date":"2025-05-08T09:36:49","date_gmt":"2025-05-08T06:36:49","guid":{"rendered":"https:\/\/sinatip.com.tr\/?p=1504"},"modified":"2025-05-08T09:47:43","modified_gmt":"2025-05-08T06:47:43","slug":"development-and-validation-of-multiplex-real-time-pcr-for-simultaneous-detection-of-six-bacterial-pathogens-causing-lower-respiratory-tract-infections-and-antimicrobial-resistance-genes","status":"publish","type":"post","link":"https:\/\/sinatip.com.tr\/tr\/development-and-validation-of-multiplex-real-time-pcr-for-simultaneous-detection-of-six-bacterial-pathogens-causing-lower-respiratory-tract-infections-and-antimicrobial-resistance-genes\/","title":{"rendered":"Alt solunum yolu enfeksiyonlar\u0131na neden olan alt\u0131 bakteriyel patojenin ve antimikrobiyal diren\u00e7 genlerinin e\u015f zamanl\u0131 tespiti i\u00e7in multipleks ger\u00e7ek zamanl\u0131 PCR&#8217;nin geli\u015ftirilmesi ve do\u011frulanmas\u0131"},"content":{"rendered":"\n<h3 class=\"wp-block-heading\">Arka Plan<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, Streptococcus pneumoniae ve Staphylococcus aureus, k\u00fcresel olarak alt solunum yolu enfeksiyonlar\u0131n\u0131n (ASYE) ba\u015fl\u0131ca bakteriyel nedenleridir ve \u00f6nemli morbidite ve mortaliteye yol a\u00e7ar. Bu patojenlerde antimikrobiyal direncin (AMR) h\u0131zla artmas\u0131, etkili antibiyotik tedavileri i\u00e7in \u00f6nemli zorluklar olu\u015fturmaktad\u0131r. D\u00fc\u015f\u00fck kaynakl\u0131 ortamlarda, ASYE&#8217;li hastalara k\u00fclt\u00fcr sonu\u00e7lar\u0131 i\u00e7in birka\u00e7 g\u00fcn beklerken ampirik olarak antibiyotik re\u00e7ete edilir. H\u0131zl\u0131 patojen ve AMR gen tespiti, optimum antibiyotik kullan\u0131m\u0131n\u0131 te\u015fvik edebilir ve sonu\u00e7lar\u0131 iyile\u015ftirebilir.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Y\u00f6ntemler<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Burada, alt\u0131 ana patojeni ve on d\u00f6rt AMR genini do\u011frudan solunum \u00f6rneklerinden h\u0131zla tan\u0131mlamak i\u00e7in EvaGreen boyas\u0131 ve erime e\u011frisi analizi kullanarak multipleks kantitatif ger\u00e7ek zamanl\u0131 PCR geli\u015ftirdik. Patojen tespiti i\u00e7in ger\u00e7ek zamanl\u0131 PCR analizlerinin tekrarlanabilirli\u011fi, do\u011frusall\u0131\u011f\u0131, tespit s\u0131n\u0131r\u0131 (LOD), DNA kontrol kar\u0131\u015f\u0131mlar\u0131 ve sivriltilmi\u015f trakeal aspirat kullan\u0131larak de\u011ferlendirildi. RT-PCR analizlerinin performans\u0131 daha sonra yo\u011fun bak\u0131m hastalar\u0131n\u0131n 50 trakeal aspirat ve balgam \u00f6rne\u011finde alt\u0131n standart, konvansiyonel k\u00fclt\u00fcrle kar\u015f\u0131la\u015ft\u0131r\u0131ld\u0131.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Sonu\u00e7lar<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">RT-PCR analizlerinin duyarl\u0131l\u0131\u011f\u0131 K. pneumoniae, A. baumannii, P. aeruginosa, E. coli i\u00e7in %100 ve S. aureus i\u00e7in %63,6 idi ve \u00f6zg\u00fcll\u00fck %87,5 ile %97,6 aras\u0131nda de\u011fi\u015fiyordu. \u0130ki y\u00f6ntem aras\u0131ndaki t\u00fcm patojenlerin kappa korelasyon de\u011ferleri 0,63 ile 0,95 aras\u0131nda de\u011fi\u015fiyordu. Hedef bakterilerin tespit s\u0131n\u0131r\u0131 1600 CFU\/ml idi. PCR analizlerinden elde edilen kantitatif sonu\u00e7lar, trakeal aspiratlar\u0131n kantitatif k\u00fclt\u00fcr\u00fcyle %100 uyum g\u00f6sterdi. K\u00fclt\u00fcrle kar\u015f\u0131la\u015ft\u0131r\u0131ld\u0131\u011f\u0131nda, PCR analizleri karma enfeksiyonlar\u0131 ve S. pneumoniae&#8217;yi tespit etmede daha y\u00fcksek duyarl\u0131l\u0131k g\u00f6sterdi. Tek enfeksiyonlarda AMR geni ve AMR fenotipi tespiti aras\u0131nda y\u00fcksek d\u00fczeyde bir uyum vard\u0131.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Sonu\u00e7lar<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Multiplex kantitatif RT-PCR analizlerimiz h\u0131zl\u0131 ve basittir, ancak alt\u0131 bakteriyel LRTI patojenini ve bunlar\u0131n antimikrobiyal diren\u00e7 genlerini tespit etmede hassas ve spesifiktir ve klinik fayda a\u00e7\u0131s\u0131ndan daha fazla de\u011ferlendirilmelidir.<\/p>\n\n\n\n<div data-wp-interactive=\"core\/file\" class=\"wp-block-file\"><object data-wp-bind--hidden=\"!state.hasPdfPreview\" hidden class=\"wp-block-file__embed\" data=\"https:\/\/sinatip.com.tr\/wp-content\/uploads\/2025\/05\/article-3.pdf\" type=\"application\/pdf\" style=\"width:100%;height:600px\" aria-label=\"Alt solunum yolu enfeksiyonlar\u0131na neden olan alt\u0131 bakteriyel patojenin ve antimikrobiyal diren\u00e7 genlerinin e\u015f zamanl\u0131 tespiti i\u00e7in multipleks ger\u00e7ek zamanl\u0131 PCR&apos;nin geli\u015ftirilmesi ve do\u011frulanmas\u0131 g\u00f6m\u00fcs\u00fc.\"><\/object><a id=\"wp-block-file--media-178b6a90-fe84-42db-b827-c85fdc818783\" href=\"https:\/\/sinatip.com.tr\/wp-content\/uploads\/2025\/05\/article-3.pdf\">Alt solunum yolu enfeksiyonlar\u0131na neden olan alt\u0131 bakteriyel patojenin ve antimikrobiyal diren\u00e7 genlerinin e\u015f zamanl\u0131 tespiti i\u00e7in multipleks ger\u00e7ek zamanl\u0131 PCR&#8217;nin geli\u015ftirilmesi ve do\u011frulanmas\u0131<\/a><a href=\"https:\/\/sinatip.com.tr\/wp-content\/uploads\/2025\/05\/article-3.pdf\" class=\"wp-block-file__button wp-element-button\" download aria-describedby=\"wp-block-file--media-178b6a90-fe84-42db-b827-c85fdc818783\">indirmek<\/a><\/div>\n\n\n\n<p class=\"wp-block-paragraph\"><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Arka Plan Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, Streptococcus pneumoniae ve Staphylococcus aureus, k\u00fcresel olarak alt solunum yolu<\/p>\n","protected":false},"author":1,"featured_media":1505,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-1504","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-uncategorized-tr"],"_links":{"self":[{"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/posts\/1504","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/comments?post=1504"}],"version-history":[{"count":3,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/posts\/1504\/revisions"}],"predecessor-version":[{"id":1521,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/posts\/1504\/revisions\/1521"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/media\/1505"}],"wp:attachment":[{"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/media?parent=1504"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/categories?post=1504"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sinatip.com.tr\/tr\/wp-json\/wp\/v2\/tags?post=1504"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}