Loop-Mediated Isothermal Amplification untuk Mendeteksi Gen blaTEM sebagai Penyandi Extended-Spectrum Beta-Lactamase pada Isolat Enterobacteriaceae

Bayu A. P. Wilopo, Sunarjati Sudigdoadi, Edhyana Sahiratmadja, Intan M. W. Dewi

Abstract


Extended-spectrum beta-lactamase (ESBL) adalah suatu jenis enzim beta-laktamase yang mampu menghidrolisis penisilin, sefalosprin, dan monobaktam yang dapat dihambat oleh asam klavulanat. Enzim ini disandi oleh banyak gen, salah satunya adalah blaTEM. Untuk  mengamplifikasi gen blaTEM  selain digunakan metode polymerase chain reaction (PCR) dapat pula dilakukan metode loop-mediated isothermal amplification (LAMP) yang membutuhkan peralatan lebih sederhana dengan prosedur yang cepat dan pembacaan hasil yang  lebih mudah. Penelitian ini merupakan uji diagnostik yang bertujuan menilai sensitivitas dan spesifisitas metode LAMP serta melihat kesesuaian hasil antara metode LAMP dan metode PCR dalam mendeteksi gen blaTEM. Sebanyak 92 isolat Enterobacteriaceae diperiksa dengan metode PCR yang kemudian dibandingkan dengan metode LAMP. Didapatkan bahwa metode LAMP memiliki sensitivitas 91,4% dan spesifisitas 91,2%, serta nilai kesesuaian (kappa) sebesar 85,4%. Sebagai simpulan, metode LAMP memiliki validitas yang baik dan kesesuaian yang sangat baik dibanding dengan metode PCR. Oleh karena itu, metode LAMP dapat dijadikan pemeriksaan alternatif dalam mendeteksi gen blaTEM terutama di daerah dengan infrastruktur laboratorium terbatas.

Kata kunci:  blaTEM, Enterobacteriaceae, ESBL, LAMP, PCR

 

Loop-Mediated Isothermal Amplification for Detecting blaTEM Gene that Encodes Extended-Spectrum Beta-Lactamase in Enterobacteriaceae Isolates


Abstract

Extended-spectrum beta-lactamase (ESBL) is a beta-lactamase enzyme that is capable of hydrolyzing penicillin, cephalosporin, and monobactam, and can be inhibited by clavulanic acid. This enzyme is encoded by multiple genes, one of them is blaTEM. Polymerase chain reaction (PCR) is one of the DNA amplification methods that are frequently used; however, there are other methods that can be used including, among others, loop-mediated isothermal amplification (LAMP). LAMP requires simple equipment with quicker and easy-to-read results compared to PCR. This study was a diagnostic test to explore the sensitivity and specificity of LAMP method compared to PCR in detecting blaTEM gene. Furthermore, the concordance between LAMP and PCR methods was assessed. A total of 92 Enterobacteriaceae isolates were examined by PCR and LAMP methods and compared. The result showed that the LAMP method had a sensitivity of 91.4% and a specificity of 91.2% with a concordance value (kappa) of 85.4%. In conclusion, LAMP method has a good validity and a very good conformity compared to the PCR method. Therefore, LAMP method can be used as an alternative diagnostic test, especially in limited settings. [MKB. 2015;47(4):242–9]

Key words: blaTEM, Enterobacteriaceae, ESBL, LAMP, PCR

 

DOI: 10.15395/mkb.v47n4.618


Keywords


blaTEM, Enterobacteriaceae, ESBL, LAMP, PCR

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