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Journal : Medical Journal of Indonesia

Optimizing real-time PCR method to detect Leptospira spp. in human blood and urine specimens Karuniawati, Anis; Yasmon, Andi; Ningsih, Ika
Medical Journal of Indonesia Vol 21, No 1 (2012): February
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (925.644 KB) | DOI: 10.13181/mji.v21i1.472

Abstract

Background: Leptospirosis is an acute infectious disease in humans caused by Leptospira spp. and classified as a zoonosis. Clinical symptoms of leptospirosis are nonspecific and the current available laboratory method for detecting Leptospira spp. is difficult, which resulted to the misdiagnosis of this disease. Therefore, the rapid and accurate method is needed to diagnose the disease. This study was aimed to optimize molecular diagnostic test using real-time PCR assay as a rapid, sensitive and specific method for the detection of pathogenic Leptospira spp. in humans.Methods: Bacterial DNA was extracted by DNA extraction kit according to the manufacturer’s instructions. Primers and probes used in this study was based on previous and published research. The assay is performed using PCR-IQTM5, iCycler Multicolor real-time PCR detection system. Specificity of the primer used was evaluated towards some bacterial pathogens.Results: Limit detection of the DNA was 0.375 fg/ml and the primers used does not cross-react with the genomes of the pathogens tested. Limit detection of DNA in blood is 150 fg/μl, and in urine is 1470 fg/μl.Conclusion: Real-time PCR test is a rapid and accurate method for detecting pathogenic Leptospira spp. in human specimens. Further research is needed to determine the sensitivity and specificity of real-time PCR tests compared with other diagnostic methods in clinical settings. (Med J Indones 2012;21:13-7)Keywords: Leptospirosis, Leptospira, optimization, real-time PCR
Genotypic pattern of fluoroquinolone resistance among extended-spectrum beta-lactamase-producing Escherichia coli Febrianti, Tati; Puspandasari, Nelly; Febriyana, Dwi; Weaver, Tom; Karuniawati, Anis
Medical Journal of Indonesia Vol. 33 No. 4 (2024): December
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13181/mji.oa.247544

Abstract

BACKGROUND Fluoroquinolone (FQ) is one of the therapeutic options for treating extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESBL-Ec) infection, but its use could increase the resistance level of ESBL-Ec. This study aimed to analyze the resistant genes responsible for ESBL production and FQ resistance among the ESBL-Ec isolated from healthy humans and communal wastewater. METHODS This was an observational study using stored isolates and laboratory data. Genome sequencing was done on 43 E. coli DNA isolates before resistance genes, mutations, and high-risk clones were examined through bioinformatic data analysis. RESULTS The analysis of 39 ESBL-Ec isolates showed ESBL genes, including blaCTX-M-55 (56%), blaCTX-M-15 (31%), and blaCTX-M-27 (8%). ESBL-Ec isolates exhibited mutations in gyrA (54%), gyrB (0%), parC (28%), and parE (10%). Plasmid-mediated quinolone resistance genes detected included qnrS1 (54%), qnrS13 (13%), qnrB2 (3%), aac(6’)-Ib-cr5 (3%), qepA (0%), and oqxAB (0%). ESBL and FQ resistance genes were simultaneously detected in 33 ESBL-Ec isolates, with high-risk clones identified as ST155, ST10, ST23, ST38, ST131, and ST69 Cplx. CONCLUSIONS ESBL and FQ resistance genes were simultaneously detected in ESBL-Ec isolated from healthy humans and communal wastewater.
Co-Authors . Andriansjah Ade Dharmawan Amalia Almira Amarila Malik Amin Soebandrio Andi Annisa Rusyda Khafiyani Andi Yasmon Andra Aswar Andra Aswar, Andra Angky Budianti Anwar S. Ibrahim Aprilia Rakhmawati Ardiana Kusumaningrum Ari Fahrial Syam Ari Prayitno Aria Kekalih Aryo Tedjo Asmarani Kusumawati Asmarani Kusumawati Augustine Natasha Aulia Rizki Bachti Alisyahbana Badriul Hegar Badriul Hegar Bambang Madiyono Burhanuddin Iskandar Ceva W. Pitoyo Chairunissa T. Rizal Cleopas Martin Rumende Cleopas Martin Rumende Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Dimas Seto Prasetyo Dwiana Ocviyanti E. Risdiyani Em Yunir Em Yunir, Em Erwin Christian Susanto Esthika Dewiasty Esthika Dewiasty, Esthika Falasiva, Rezyta Fariz Nurwidya Febrianti, Tati Febriyana, Dwi Futihati Ruhama Zulfa Gestina Aliska Gortap Sihotang Hana Paraswati Putri Handaya, Adeodatus Y. Hindra Irawan Satari Hindra Irawan Satari Ida Parwati Ika Ningsih Indah S. Widyahening Iswanto, J. Joyce Bratanata Joyce Bratanata, Joyce Kambang Sariadji Khie Chen Kuntjoro Harimurti Lalisang, Toar JM. Linosefa Linosefa Manuel Lamberto Willem Mboeik Martin Hartiningsih Mboeik, Manuel Lamberto Willem Moehario, Lucky Hartati Muhammad K Azwar, Muhammad K Mulya Rahma Karyanti, Mulya Rahma Mulyadi M. Djer Muziasari, Windi Nabila Maudy Salma Nasution, Safruddin Nelly Puspandari Nie Nie Nina Dwi Putri Nuraini I Susanti Permatasari, Tri Wijayanti Pratama Wicaksana Pratiwi Pujilestari Sudarmono Pratiwi Sudharmono Pratiwi Sudharmono, Pratiwi Prawoto Prawoto Prawoto Prawoto Pujilestari, Ratih Purwantyastuti Purwantyastuti Puspandasari, Nelly Rakhmawati, Aprilia Resti H. Lestari Reynaldo Reynaldo Reynaldo Reynaldo, Reynaldo Rezyta Falasiva Riamin Sitorus Rianto Setiabudi, Rianto Rida Tiffarent Rosdiana Irawati Rudyanto Sedono Rudyanto Sedono Rudyanto Sedono, Rudyanto S. Nilawati Santya Fatma Dewi Sari Wiraswasty Saunar, Rofy S. Shindy Claudya Aprianti Simanjuntak, Glory Gelarich Sitorus, Truly Panca SJATHA, FITHRIYAH Subari A. Riyanto Sudigdo Sastroasmoro Sukman T. Putra Sunarno Sunarno Susan M. Noor Susan M. Noor, Susan Susan Maphilindawati Noor Susanti, Nuraini I Susanti, Nuraini I Susanto, Erwin Christian Sweety Pribadi SYADZA RHIZKY PUTRI AKHMAD T. M. Sudiro Teguh Harjono Karjadi Teguh Harjono Karjadi, Teguh Harjono TETSUHIRO MATSUZAWA Thomas Robertus Tjampakasari, Conny Riana TOHRU GONOI Tony Loho Trisni U Dewi, Trisni U Truely Panca Sitorus Usman, Nurhayat Weaver, Tom Wia Melia Windi Muziasari Yefta Moenadjat Yeva Rosana Yulia R. Saharman Yulia Rosa Saharman