Une plaque d’alopécie peu commune Le Govic, Y.; Miossec, C.; Courdurié, C. ...
Journal de mycologie médicale,
12/2014, Letnik:
24, Številka:
4
Journal Article
Microscopy is the reference standard for routine laboratory diagnosis in faecal parasitology but there is growing interest in alternative methods to overcome the limitations of microscopic ...examination, which is time-consuming and highly dependent on an operator's skills and expertise. Compared with microscopy, DNA detection by PCR is simple and can offer a better turnaround time. However, PCR performances remain difficult to assess as most studies have been conducted on a limited number of positive clinical samples and used in-house PCR methods. Our aim was to evaluate a new multiplex PCR assay (G-DiaParaTrio; Diagenode Diagnostics), targeting Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica. To minimize the turnaround time, PCR was coupled with automated DNA extraction (QiaSymphony; Qiagen). The PCR assay was evaluated using a reference panel of 185 samples established by routine microscopic examination using a standardized protocol including Ziehl–Neelsen staining and adhesin detection by ELISA (E. histolytica II; TechLab). This panel, collected from 12 French parasitology laboratories, included 135 positive samples for G. intestinalis (n = 38), C. parvum/C. hominis (n = 26), E. histolytica (n = 5), 21 other gastrointestinal parasites, together with 50 negative samples. In all, the G-DiaParaTrio multiplex PCR assay identified 38 G. intestinalis, 25 C. parvum/C. hominis and five E. histolytica leading to sensitivity/specificity of 92%/100%, 96%/100% and 100%/100% for G. intestinalis, C. parvum/C. hominis and E. histolytica, respectively. This new multiplex PCR assay offers fast and reliable results, similar to microscopy-driven diagnosis for the detection of these gastrointestinal protozoa, allowing its implementation in routine clinical practice.
A 36-year-old woman returning from holiday in Portugal presented with bilateral ocular pain and acute visual loss. Eye examination revealed an inflammation in the anterior segment associated with ...corneal keratitis. During her trip, she wore contact lenses and rinsed the storage case using tap water. Corneal scrapings, contact lenses and cleaning solution from the case were inoculated onto agar plates supplemented with heat-inactivated bacteria. Bilateral Acanthamoeba keratitis was diagnosed, as microscopic examination of cultures from the last two samples revealed typical, spherical double-walled cystic structures (Fig. 1).
Besides the potential to identify a wide variety of gastrointestinal parasites, microscopy remains the reference standard in clinical microbiology for amoeba species identification and, especially ...when coupled with adhesin detection, to discriminate the pathogenic Entamoeba histolytica from its sister but non-pathogenic species Entamoeba dispar/Entamoeba moshkovskii. However, this approach is time-consuming, requires a high-level of expertise that can be jeopardized considering the low prevalence of gastrointestinal parasites in non-endemic countries. Here, we evaluated the CE-IVD-marked multiplex PCR (ParaGENIE G-Amoeba, Ademtech) targeting E. histolytica and E. dispar/E. moshkovskii and Giardia intestinalis.
This evaluation was performed blindly on a reference panel of 172 clinical stool samples collected prospectively from 12 laboratories and analysed using a standardized protocol relying on microscopy (and adhesin detection by ELISA for the detection of E. histolytica) including G. intestinalis (n = 37), various amoeba species (n = 55) including E. dispar (n = 15), E. histolytica (n = 5), as well as 17 other gastrointestinal parasites (n = 80), and negative samples (n = 37).
This new multiplex PCR assay offers fast and reliable results with appropriate sensitivity and specificity for the detection of G. intestinalis and E. dispar/E. moshkovskii from stools (89.7%/96.9% and 95%/100%, respectively). Detection rate and specificity were greatly improved by the PCR assay, highlighting several samples misidentified by microscopy, including false-negative and false-positive results for both E. dispar/E. moshkovskii and E. histolytica.
Given the clinical relevance of amoeba species identification, microbiologists should be aware of the limitations of using an algorithm relying on microscopy coupled with adhesin detection by ELISA.
Twenty-five patients, of whom 22 were renal transplant recipients, developed Pneumocystis jirovecii infections at the nephrology department of Reims University Hospital (France) from September 2008 ...to October 2009, whereas only four sporadic cases had been diagnosed in this department over the 14 previous years.
This outbreak was investigated by analysing patient encounters and P. jirovecii types.
A transmission map was drawn up. P. jirovecii typing at DHPS, ITS and mtLSU rRNA sequences was performed in the patients of the cluster (18 patients with Pneumocystis pneumonia (PCP) and seven colonized patients), 10 unlinked control patients (six PCP patients and four colonized patients), as well as 23 other patients diagnosed with P. jirovecii (nine PCP patients and 14 colonized patients) in the same department over a three-year post-epidemic period.
Eleven encounters between patients harbouring the same types were observed. Three PCP patients and one colonized patient were considered as possible index cases. The most frequent types in the cluster group and the control group were identical. However, their frequency was significantly higher in the first than in the second group (P < 0.01). Identical types were also identified in the post-epidemic group, suggesting a second outbreak due to the same strain, contemporary to a disruption in prevention measures.
These results provide additional data on the role of both PCP and colonized patients as infectious sources. Longitudinal screening of P. jirovecii types in infected patients, including colonized patients, is required in the investigation of the fungus's circulation within hospitals.
Fungal otitis (otomycosis) is a common infection encountered by otolaryngologists. Nevertheless, its management can be challenging because of its high recurrence rate and of the limited therapeutic ...options. A 45-year-old woman suffered from recurrent otomycosis. The ineffectiveness of successive antibiotic cures and repeated topical treatments with nystatin and then with econazole cream led to perform microbiological analyses. Culture of ear swab grew Aspergillus niger. The use of a 1% voriconazole sterile solution previously validated for treatment of eye infections was considered after ensuring the absence of known ototoxic effects of the antifungal and of the excipients. The patient was advised to apply locally this voriconazole solution daily for 14 days (3 drops, 3–4 times a day). Full recovery was obtained at the end of the treatment, and no relevant side effects were noticed. More than one year after completion of therapy, there was no recurrence. Our observation shows that voriconazole 1% solution is an interesting option for treating otomycosis which failed to respond to usual therapeutic options. Further prospective studies are now warranted to confirm these findings.
In vitro susceptibility of 933 Candida isolates, from 16 French hospitals, to micafungin was determined using the Etest in each center. All isolates were then sent to a single center for ...determination of MICs by the EUCAST reference method. Overall essential agreement between the two tests was 98.5% at ±2 log2 dilutions and 90.2% at ±1 log2 dilutions. Categorical agreement was 98.2%. The Etest is a valuable alternative to EUCAST for the routine determination of micafungin MICs in medical mycology laboratories.
Cryptosporidiosis is an important though underreported public health concern. Molecular tools might be helpful in improving its diagnosis. In this study, ZR Fecal DNA MiniPrep™ Kit (ZR) and ...NucliSens® easyMAG® (EM) were compared using four
Cryptosporidium
-seeded feces and 29
Cryptosporidium
-positive stools. Thereafter, ZR was selected for prospective evaluation of
Cryptosporidium
detection by 18S rDNA and LAXER quantitative PCR (qPCR) in 69 stools from 56 patients after
Cryptosporidium
detection by glycerin, modified Ziehl–Neelsen (ZN) and auramine–phenol (AP) stainings. The combination of any of the two extraction methods with 18S qPCR yielded adequate detection of
Cryptosporidium
in seeded stools, but the ZR kit showed the best performance. All 29
Cryptosporidium
-positive samples were positive with 18S qPCR, after both ZR and EM extraction. However, false-negative results were found with LAXER qPCR or nested PCR. Cryptosporidiosis was diagnosed in 7/56 patients. All the microscopic methods enabled the initial diagnosis, but
Cryptosporidium
was detected in 12, 13, and 14 samples from these seven patients after glycerin, ZN, and AP staining respectively. Among these samples, 14 and 12 were positive with 18S and LAXER qPCR respectively. In two patients,
Cryptosporidium
DNA loads were found to be correlated with clinical evolution. Although little known, glycerin is a sensitive method for the initial detection of
Cryptosporidium
. When combined with 18S qPCR, ZR extraction, which had not been evaluated so far for
Cryptosporidium
, was an accurate tool for detecting
Cryptosporidium
and estimating the oocyst shedding in the course of infection.
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