| 1. |
- Edberg, Andreas, et al.
(författare)
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A comparative study of three different PCR assays for detection of Mycoplasma genitalium in urogenital specimens from men and women
- 2008
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Ingår i: Journal of Medical Microbiology. - : Microbiology Society. - 0022-2615 .- 1473-5644. ; 57:Pt 3, s. 304-309
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Tidskriftsartikel (refereegranskat)abstract
- The aim of this study was to compare conventional 16S rRNA gene PCR, real-time 16S rRNA gene PCR and real-time Mycoplasma genitalium adhesin protein (MgPa) gene PCR as detection methods for M. genitalium infection. The study also determined the prevalence of M. genitalium in male and female patients attending a sexually transmitted infections clinic in a rural area in the west of Sweden. First void urine (FVU) and/or urethral swabs were collected from 381 men, and FVU and/or cervical swabs and/or urethral swabs were collected from 298 women. A total of 213 specimens were used in the PCR comparative study: 98 consecutively sampled specimens from patients enrolled in the prevalence study, 36 consecutively sampled specimens from patients with symptoms of urethritis and 79 specimens from patients positive for M. genitalium by real-time MgPa gene PCR in the prevalence study. A true-positive M. genitalium DNA specimen was defined as either a specimen positive in any two PCR assays or a specimen whose PCR product was verified by DNA sequencing. The prevalence of M. genitalium infection in men and women was 27/381 (7.1 %) and 23/298 (7.7 %), respectively. In the PCR comparative study, M. genitalium DNA was detected in 61/76 (80.3 %) of true-positive specimens by conventional 16S rRNA gene PCR, in 52/76 (68.4 %) by real-time 16S rRNA gene PCR and in 74/76 (97.4 %) by real-time MgPa gene PCR. Real-time MgPa gene PCR thus had higher sensitivity compared with conventional 16S rRNA gene PCR and had considerably increased sensitivity compared with real-time 16S rRNA gene PCR for detection of M. genitalium DNA. Real-time MgPa gene PCR is well suited for the clinical diagnosis of M. genitalium.
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| 2. |
- Hjorth, SV, et al.
(författare)
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Sequence-based typing of Mycoplasma genitalium reveals sexual transmission
- 2006
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Ingår i: Journal of Clinical Microbiology. - 0095-1137 .- 1098-660X. ; 44:6, s. 2078-2083
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Tidskriftsartikel (refereegranskat)abstract
- Mycoplasma genitalium causes male nonchlamydial, nongonococcal urethritis and is associated with cervicitis and pelvic inflammatory disease in women. Epidemiological studies indicate that M. genitalium is sexually transmitted, and the aim of the present study was to further substantiate this by means of a DNA typing system. A typing assay based on a diagnostic mgpB gene PCR was developed, evaluated, and applied directly to urogenital specimens. The assay had a low limit of detection and hence a high typeability. Sequences of isolates from 52 unrelated patients were divided into 29 different sequence types, giving a discriminatory index of 0.95. Two to six M. genitalium-positive specimens were collected from each of 44 patients over a median interval of 56 days (range, 11 to 1,395). Forty had the same sequence type in consecutive specimens. Specimens collected from two men were repeatedly positive at intervals of 472 and 1,395 days, respectively, but the sequence types had changed. A new strain was introduced in one sexual dyad, and the sequence types changed subsequently. Seventy-nine M. genitalium-positive specimens from 19 couples were investigated, and all partners initially had concordant sequence types, but one couple had discordant types at one time point before a newly introduced strain took over. The present typing system is simple and reproducible and has an excellent discriminatory capacity which might prove useful in studies of sexual networks and for evaluation of treatment failures. In the laboratory, this system may document the uniqueness of newly isolated M. genitalium strains.
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| 3. |
- Jurstrand, Margaretha, et al.
(författare)
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Detection of Mycoplasma genitalium in urogenital specimens by real-time PCR and by conventional PCR assay
- 2005
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Ingår i: Journal of Medical Microbiology. - : Microbiology Society. - 0022-2615 .- 1473-5644. ; 54:1, s. 23-29
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Tidskriftsartikel (refereegranskat)abstract
- A real-time LightCycler PCR (LC-PCR) with hybridization probesfor detection of Mycoplasma genitalium in endocervical and firstvoid urine specimens was developed and compared to a conventionalPCR. The primers for both assays were identical and designedto amplify a 427 bp fragment of the 16S rRNA gene of M. genitalium.The LC-PCR assay had a detection limit of < 5 bacterial genomesper reaction when dilutions of genomic DNA from a type strainof M. genitalium were tested. First void urine from 398 menand first void urine and endocervical specimens from 301 womenattending an STD clinic were analysed by LC-PCR and by the conventionalPCR. Using the conventional PCR as reference, the LC-PCR hada specificity of 99.7 % and a sensitivity of 72.2 % for thedetection of M. genitalium in first void urine samples frommen. There was no significant difference in the performanceof the LC-PCR assay compared to the conventional PCR when endocervicalswabs were considered (58 and 65 %, respectively) or with aset of endocervical swab/urine specimens for which the LC-PCRassay detected 73 % of the infections (specificity = 98.6 %and sensitivity = 68.2 %) while the conventional PCR detected85 % of the infections. With female urine specimens there wasa significant difference between the two assays (38 and 73 %,respectively; P = 0.01 McNemar's test). This illustrates theneed to analyse both endocervical and urine specimens, becauseM. genitalium DNA was detected in only one of the two specimensin a great number of the M. genitalium-infected women. The lowersensitivity of the LC-PCR assay was probably caused by a combinationof inhibition and limitations regarding the amount of templateDNA. The LC-PCR assay was easy to perform and the simultaneousamplification and detection eliminated the need for furtherhandling of PCR products. With improvement in sample preparationmethods and increased volumes of the template DNA, the LC-PCRassay could be a useful routine diagnostic method.
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| 4. |
- Mellenius, Harriet, et al.
(författare)
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[Mycoplasma genitalium should be suspected in unspecific urethritis and cervicitis. A study from Vasterbotten confirms the high prevalence of the bacteria]
- 2005
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Ingår i: Läkartidningen. - 0023-7205 .- 1652-7518. ; 102:47, s. 3538-3541
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Tidskriftsartikel (refereegranskat)abstract
- The microbe Mycoplasma genitalium has in several studies been proposed as an individual cause of non-gonococcal urethritis (NGU) in men, and has been associated with pelvic inflammatory disease (PID) and salpingitis. The prevalence of M genitalium has generally been 50-90% of the prevalence of C trachomatis, and this seems to be the case in Sweden as well. This is the first study of the pathogenesis and prevalence of M genitalium in northern Sweden. In total 823 samples, 340 from women and 483 from men, were screened for M genitalium by using a PCR method. Thirtythree (4.0%) patients, 13 (3.8%) women and 20 (4.1%) men, were infected by M genitalium. In the same group 60 (7.3%) patients, 16 (4.7%) women and 44 (9.1%) men, were infected by Chlamydia trachomatis. None of the 22 patients that were tested after treatment with azitromycin was still infected.
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