Identification of clinical strains of Candida albicans by DNA fingerprinting with the polymerase chain reaction

Abstract

Summary. DNA polymorphisms generated by the polymerase chain reaction (PCR) were used to differentiate clinical isolates of Candida. This PCR method employed single primers that were originally designed as hybridization probes for DNA fingerprinting experiments to probe mini‐satellite and microsatellite DNA sequences. To evaluate this procedure, 35 isolates from 20 patients in several intensive care units and 12 isolates obtained from the oral cavities of healthy dental patients were fingerprinted. The PCR‐fingerprint patterns of isolates of Candida albicans from the immunocompromised patients revealed fewer differences than isolates from the dental service. Multiple isolates from different body sites of the same patients revealed that patients may harbour isolates of Candida with the same or different PCR‐fingerprints. Since this method is generally simpler and faster than established methods of biotyping medically important yeasts, PCR‐fingerprinting may prove useful for the surveying of large numbers of pathogens for epidemi‐ological studies. Zusammenfassung. Mittels PCR erhaltene DNA‐Polymorphismen wurden zur Differen‐zierung von klinischen Candida‐Isolaten einge‐setzt. Einzelne Primer, welche spezifisch Mini‐ und Mikrosatelliten‐DNA‐Sequenzen amplifizieren, wurden fur das PCR‐Fingerprinting genutzt. Mit dieser Methode wurden 35 Isolate, welche von 20 Patienten ver‐schiedener Intensivtherapiestationen stammten, sowie 12 Isolate aus der Mundhöhle gesunder stomatologischer Patienten untersucht. Die PCR‐Fingerprint‐Muster der Candida‐Stämme, die von immunsupprimierten Patienten isoliert wurden, zeigten dabei geringere Unterschiede als die der Oralisolate. Isolate von verschiedenen Körperstellen des gleichen Patienten ergaben entweder identische oder differierende PCR‐Fingerprints. Die PCR ist in vielen Fällen leichter und schneller durchführbar als andere, bereits etablierte, genetische Verfahren für die Typisierung von Krankheitserregern. Für die Untersuchung einer großen Anzahl von Isolaten im Rahmen epidemiologischer Studien ist diese Methode deshalb von besonderem Wert.

One Small Step for a Yeast - Microevolution within Macrophages Renders Candida glabrata Hypervirulent Due to a Single Point Mutation

“…Total genomic DNA (gDNA) was recovered from fungal cells by a standard nucleic acid extraction protocol adapted from [56] , using phenol∶chloroform and glass beads in a Precellys 24 homogeniser (PeqLab, Germany). The fingerprinting PCR reaction was performed as described previously [57] . Primers used were the M13 primer ( GAGGGTGGCGGTTCT ) and a primer consisting of the repeat sequence (GACA) 4 .…”

DNA fingerprinting of Fusarium solani isolates related to a cutaneous infection in a sea turtle

“…At present, PCR techniques appear to be useful for the typing of fungal isolates [11,12]. Use of RAPDs have been helpful for the epidemiology of clinical Candida species [13,14]. Cryptococcus neoformans [15], Aspergillus fumigatus [16–19], Scedosporium prolificans [20] and other fungi [21].…”

Analysis and effect of the use of biofertilizers on <em>Trifolium rubens</em> L., a preferential attention species in Castile and Leon, Spain, with the aim of increasing the plants conservation status

“…A Random Amplification of Polymorphic DNA (RAPD) was performed to confirm that the isolated strain not clones. Genetic profiles were performed using the following PCR conditions: 12.5 µl of Dream Taq Green PCR Master Mix (Thermo Fisher Scientific), 2.5 µl of primer M13 with a concentration of 20 µM (5′-CAGGGTGGCGGTTCT-3′) [16], 8.5 µl of miliQ water and 1.5 of DNA. The parameters of the thermocycler were: 95 °C for 10 min; 35 cycles of 94 °C for 1 min, 45 °C for 1 min and 72 °C for 2 min; and a final extension of 72 °C for 7 minutes.…”