Simultaneous detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR) assay in cooling tower water samples from Jakarta, Indonesia

Yasmon, Andi; Yusmaniar, Yusmaniar; Anis, Anis; Bela, Budiman

doi:10.13181/mji.v19i4.408

Cited by 6 publications

(4 citation statements)

References 19 publications

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“…Metode duplex PCR lebih efisien, hemat waktu, dan biaya jika dibandingkan dengan single PCR atau uniplex PCR. Beberapa peneliti telah menggunakan teknik dPCR untuk deteksi patogen, Yasmon et al (2010) telah mendeteksi secara simultan Legionella species dan Legionella pneumophila dengan dPCR pada sampel air di Jakarta. Thanananta & Thanananta (2008) mendeteksi E. coli yang berasal dari air menggunakan dPCR dengan target gen lacZ dan uidA.…”

Section: Pendahuluanunclassified

DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)

Novita

Sugiani

Taukhid

et al. 2020

Jurnal Riset Akuakultur

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Koi herpesvirus (KHV) dan Aeromonas hydrophila adalah patogen yang dapat mengkoinfeksi ikan mas secara bersamaan. Penelitian ini bertujuan untuk mengembangkan metode duplex polymerase chain reaction (dPCR), deteksi simultan untuk diagnosis KHV dan bakteri Aeromonas hydrophila pada ikan mas. Dua pasang primer yang menargetkan sekuen spesifik SphI dan gen aerolisin, yang sering digunakan untuk mendeteksi KHV dan A. hydrophila dalam uji reaksi tunggal PCR dan menghasilkan target pita PCR 290 bp dan 417 bp. Hasil penelitian menunjukkan bahwa metode duplex PCR dapat mendeteksi ganda infeksi KHV dan A. hydrophila pada ikan mas dan metode ini lebih efektif mendeteksi dua patogen secara bersamaan dalam satu reaksi PCR pada suhu pradenaturasi, 94°C selama dua menit, denaturasi pada suhu 95°C selama satu menit, annealing pada suhu, 55°C selama satu menit, dan 72°C selama satu menit, dengan 30 siklus amplifikasi dan final extention pada suhu 72°C selama lima menit. Metode dPCR untuk deteksi simultan kedua patogen adalah salah satu metode yang dapat diaplikasikan untuk deteksi koinfeksi virus dan bakteri dalam satu reaksi PCR.Koi herpesvirus (KHV) and Aeromonas hydrophila are pathogens that can co-infect common carp. This study aimed to develop a duplex polymerase chain reaction (dPCR) method to detect KHV and Aeromonas hydrophila in common carp simultaneously. Two pairs of primers targeted the specific sequences of SphI and aerolysin genes, often used in detecting KHV and A. hydrophila, in a single PCR reaction test and produced target bands of PCR 290 bp and 417 bp. This proposed method was more effective in simultaneously detecting the two pathogens in one PCR reaction at pre-naturation temperature of 94°C for two minutes, denaturation at 95°C for one minute, annealing at temperature, 55°C for one minute, and 72°C for one minute, with 30 cycles of amplification and final extension at 72°C for five minutes. The findings showed that the duplex PCR method could be used to double detect KHV and A. hydrophila infection in common carp. The duplex PCR method for simultaneous detection of both pathogens is one method that can be applied for the detection of co-infection of viruses and bacteria in a PCR reaction.

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Section: Pendahuluanunclassified

DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)

Novita

Sugiani

Taukhid

et al. 2020

Jurnal Riset Akuakultur

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“…Legionella merupakan bakteri yang berkaitan dengan air dan tersebar luas di lingkungan, dan mampu bertahan dalam kondisi ekstrim suhu tinggi (Yasmon et al, 2010). Bakteri ini dapat ditemukan pada sumbersumber air alami dan juga buatan manusia.…”

Section: Pendahuluanunclassified

Untitled

Aksono¹,

Farahdiba²,

Hestianah³

2017

JVet

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ABSTRAKLegionella pneumophila adalah bakteri Gram-negatif berbentuk batang yang dapat menyebabkan penyakit nosokomial dan pneumonia. Tujuan penelitian ini adalah untuk mendeteksi keberadaan bakteri L. pneumophila pada air kolam renang di Kota Surabaya dengan menggunakan nested Polymerase Chain Reaction (PCR) berbasis gen spesifik L. pneumophila (mip gene). Penelitian ini menggunakan metode purposive sampling. Sebanyak sepuluh sampel diambil dari lima kolam renang. Sampel diambil sebanyak 200 mL dari air kolam renang di setiap lokasi. Hasil dari 10 sampel yang diuji menggunakan nested PCR, satu sampel menunjukkan hasil positif untuk L.pneumophila, dan sembilan sampel menunjukkan hasil negatif. Bakteri L. pneumophila ditemukan pada sampel air kolam dengan suhu yang lebih tinggi (>30ºC). Satu sampel positip tersebut ketika dilanjutkan terhadap analisis serogrup terlihat bahwa bakteri L. pneumophila yang terdeteksi pada air kolam renang di Kota Surabaya termasuk L. pneumophila serogrup 9 (98%) dan serogrup 10 (98%). Terdeteksinya bakteri L. pneumophila ini diharapkan dapat meningkatkan kesadaran dokter dan ahli mikrobiologi tentang penyebaran L. pneumophila dan juga bermanfaat untuk mengontrol agen Legionellosis. ABSTRACTLegionella pneumophila is a Gram-negative bacillus that causes nosocomial and community-acquired pneumonia. The aim of this research was to detect the presence of bacteria of L. pneumophila species in the swimming pools water of Surabaya city by using nested Polymerase Chain Reaction (PCR) assay of a specific gene for L. pneumophila (mip gene). This study used purposive sampling method. A total of 10 water samples were collected from five swimming pools consisting of 200 mL water for each swimming pool. The results showed that of 10 samples tested by nested PCR, one sample was positive for L. pneumophila, and nine samples were negative. L. pneumophila were found in pool water samples with a higher temperature (>30ºC).Serogrouping analysis of positive sample that L. pneumophila bacteria detected in the water sample of swimming pool in Surabaya was L. pneumophila serogroup 9 (98%) and serogroup 10 (98%). L. pneumophila detection of bacteria is expected to raise the awareness of physician and microbiologists about the transmission of L. pneumophila and will also be useful for controlling the agents.

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“…11 Research conducted by Yasmon et al from 9 cooling tower water samples taken from buildings in Jakarta, Indonesia from November 2007 to June 2008 tested with duplex PCR (dPCR) tests, 6 of them were positive for Legionella sp., 1 positive for L. pneumophila, and 2 negative for both. 12 Another study also examined Legionella bacteria where as many as 17 water samples were taken from 17 different sources, of which 10 samples were obtained from hospitals in North Jakarta, and 7 samples from West Jakarta. Water samples were all cultivated in all four media variants Culture media removed from the incubator on days 3, 7 and 10 to observe growth and presence of colonies corresponding to the characteristics of L. pneumophila.…”

Section: Introductionmentioning

confidence: 99%

Screening of Legionella pneumophila from Well Water in Magetan Regency, Indonesia

Hapsari,

Kurniawan,

Sucipto

et al. 2023

IJTID

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Legionellosis is a respiratory infection caused by Legionella pneumophila, a bacterium that can infect protozoa and human lung cells. The disease can be mild or severe, and sometimes fatal, especially in immunocompromised people. Some types of antibiotics that can be used to treat Legionella disease are macrolides such as rifampicin, azithromycin and clarithromycin, as well as fluoroquinolones such as levofloxacin and moxifloxacin. The aim was to investigate the distribution and prevalence of Legionella in well water. This study collected and tested water samples from a different well water in Magetan Regency, East Java, Indonesia using DNA extraction, two-step PCR methods, and visualization with 1.5% agarose gel in UV transluminator. The results were visualized with QGIS 3.28.6 and compared with other tests. The results showed that none of the eight water samples were contaminated with L. pneumophila compared with positive control (403 bp).

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Simultaneous detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR) assay in cooling tower water samples from Jakarta, Indonesia

Cited by 6 publications

References 19 publications

DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)

DUPLEX POLYMERASE CHAIN REACTION UNTUK DETEKSI SIMULTAN KOI HERPESVIRUS DAN Aeromonas hydrophila PADA IKAN MAS (Cyprinus carpio)

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Screening of Legionella pneumophila from Well Water in Magetan Regency, Indonesia

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