Use of restriction fragment length polymorphism as a genetic marker for typing Mycobacterium avium strains

Abstract

Restriction fragment length polymorphism (RFLP) was used to study 75 clinical isolates identified as Mycobacterium avium. Two repetitive insertion sequences, IS1311 and IS900, were used as DNA probes. Although less than 25% of isolates showed RFLP patterns with IS900, all strains gave banding patterns with IS1311. M. avium strains isolated from patients with AIDS exhibited marked polymorphism with both probes.

Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements

“…Recently, genetic markers like the RFLP typing using insertion sequences IS 1245 , IS 1311 , IS 901 , IS 902 and IS 1110 and PFGE appeared very useful to type M. avium ssp. avium isolates [262, 263, 267–277]. Serotyping and IS 1245 RFLP typing correlate only partly [263].…”

Molecular features ofMycobacterium aviumhuman isolates carrying a single copy of IS1245and IS1311per genome

“… RFLP patterns of Mycobacterium avium Mav III Colombian isolates. IS 1245 ‐ and IS 1311 ‐RFLP profiles were performed using previously described procedures (Roiz et al , 1995; van Soolingen et al , 1998) using the probes IS 1245 (left panel) and IS 1311 (right panel). Lanes 1 and 18, reference strains M. avium R13 and M. avium IW49, respectively; lanes 2–15, M. avium Mav III Colombian isolates: 32B, 37B, 38B, 43B, 44B, 45B, 47B, 48B, 49B, 50B, 51B, 53B, 54B, and 56B.…”

“…Analysis of IS 1245 ‐ and IS 1311 ‐RFLP profiles was performed using previously described procedures (Roiz et al , 1995; van Soolingen et al , 1998). All but two of the Mav III isolates (19/21) showed a single copy for each of the ISs: IS 1245 and IS 1311 (Fig.…”

“…The PCR amplifications of the selected target sequences were performed according to the procedures and primers described previously, as indicated in Table 2, as follows: IS 1245 (Guerrero et al , 1995); IS 1311 (Roiz et al , 1995); IS 901 (Turenne et al , 2006); the macrophage‐induced gene ( mig ) gene (Beggs et al , 2000); and IS 900 (Bull et al , 2003). The IS 1613 was also amplified using IS 1613 .2R and IS 1613 .1 as primers (Table 2), and the following PCR conditions: 94°C/5 min to denature the DNA, 35 cycles of 94°C/1 min, 56°C/1 min, and 72°C/min, with a final extension at 72°C/7 min.…”

“…The IS 1245 RFLP and IS 1311 RFLP analyses were performed using probes and conditions as described by van Soolingen et al (1998) and Roiz et al (1995), respectively. Probes were radiolabelled with α 32 P‐dCTP (Amersham Life Science) using the Megaprime DNA‐labelling system (Amersham Life Science).…”