scholarly journals Isolation of Clostridium difficile from faecal specimens – a comparison of chromID C. difficile agar and cycloserine-cefoxitin-fructose agar

2013 ◽  
Vol 62 (9) ◽  
pp. 1423-1427 ◽  
Author(s):  
Kerry C. Carson ◽  
Lusiana V. Boseiwaqa ◽  
Sara K. Thean ◽  
Niki F. Foster ◽  
Thomas V. Riley

The culture of toxigenic Clostridium difficile from stool specimens is still seen as the gold standard for the laboratory diagnosis of C. difficile infection (CDI). bioMérieux have released ChromID Cdiff chromogenic agar (CDIF) for the isolation and identification of C. difficile in 24 h. In this study, we compared CDIF to pre-reduced cycloserine-cefoxitin-fructose agar with sodium taurocholate (TCCFA) in the examination of glutamate dehydrogenase-positive faecal specimens that were either GeneOhm positive or negative, using direct culture or culture following alcohol shock. Direct culture on CDIF had a sensitivity of 100 % and recovery of 94 % while for TCCFA these were 87 % and 82 %, respectively. For GeneOhm-positive alcohol-shocked faecal samples, sensitivity and recovery on CDIF was similar to direct culture while on TCCFA they were about 10 % higher. For direct culture, there was a significant difference between growth on CDIF at 24 h and TCCFA at 48 h (P = 0.001) and between the two media at 48 h (P<0.001). A total of 142 strains of C. difficile were recovered in pure culture from all GeneOhm-positive samples used in this study and 11 (7.7 %) of these were A−B−CDT− and may represent mixed infections of toxigenic and non-toxigenic C. difficile. The most dominant ribotype was UK 014 (14.7 %) followed by 002 (11.9 %) and 020 (11.9 %), and 36 % of toxigenic isolates, including an A−B+CDT− strain, could not be assigned a UK ribotype. CDIF outperformed pre-reduced TCCFA by negating the need for alcohol shock treatment and by giving a time saving of 24 h in the isolation of C. difficile. CDIF plates were also more selective than TCCFA and C. difficile colonies were easy to identify and subculture prior to strain typing.

BioMedica ◽  
2020 ◽  
Vol 36 (4) ◽  
pp. 362-366
Author(s):  
Dr. Majid Rauf Ahmad ◽  
Dr. Iffat Javed ◽  
Dr. Sohaila Mushtaq ◽  
Dr. Rubeena Hafeez ◽  
Dr. Kanwal Hassan Cheema

Background and Objective: Dermatophyte infections require laboratory diagnosis before treatment is started. Although direct microscopy is routinely performed but culture of dermatophytes is the gold standard. However, it takes about 4 weeks for species identification on primary media. Our aim was to compare dermatophyte test medium (DTM) as a screening medium for the isolation of dermatophytes in comparison with sabouraud dextrose agar (SDA). Methods: It was a comparative study carried out at the Department of Microbiology of Post Graduate Medical Institute, Lahore over a period of nine months. Samples were collected from one hundred patients with clinically suspected dermatophytoses after taking informed written consent. The samples were examined microscopically and then inoculated on two types of culture media, one Sabouraud dextrose agar (SDA) with added chloramphenicol, gentacin and cycloheximide and other dermatophyte test medium (DTM) with added chlortetracycline, gentacin and cyclohexamide. Results: Fungal growth was observed in fifty-six samples on culture. Out of the fifty-six positive on cultures, nineteen were that of dermatophytes. Out of n = 100 patients, ten were positive on SDA while n = 14 dermatophyte species were able to grow on DTM. A significantly higher positivity (P ³ 0.05) for isolating dermatophytes was observed by DTM as compared to SDA. DTM was able to isolate (71%) of the dermatophytes in first 10 days. Isolation rate of dermatophyte species was higher (73.68%) on DTM as compared to SDA which was 52.6%. Conclusion: Authors recommend the use of dermatophyte test medium for the primary isolation and identification of dermatophyte species to be more effective and time saving.


Author(s):  
Mool Raj ◽  
Tanishu Mahajan

The study aims to assess the extent of use and adequacy of using the bio-enabled attendance system in educational institutions to develop punctuality among teachers. A fingerprint-based attendance monitoring system was used for this purpose. The sample comprises 70 teachers from an institution where a bio-enabled attendance machine was being used. The investigators collected teachers’ opinions towards using a bio-enabled attendance system for improving their punctuality with the help of a self-prepared questionnaire. The findings reveal that there is a significant difference in the teachers’ opinions towards the use of a bio-enabled attendance system with regards to their residential background and medium of teaching. There is no significant difference in the opinion of the teachers towards the use of a bio-enabled attendance system concerning their gender, computer training and teaching subjects. The results also show that the bio-enabled attendance system contributes towards improving the punctuality of teachers as it is effective, accurate, time-saving and checks the proxy system for marking the attendance.


Author(s):  
E.A. Omudu ◽  
E.U. Amuta

Domestic environmental pollution resulting from urban livestock farming was investigated in Makurdi using parasitological techniques. The test tube flotation technique was used for the parasitological analysis of animal faecal matter and soil samples collected from residential premises. Ectoparasitic fauna of dogs, goats, sheep and cattle cohabiting with humans within the same residential compound were also collected and identified. The hand-picking and body brushing methods were employed to search for ticks, fleas, lice and mites. Of the 150 soil samples examined, 55 (36.7 %) were positive for 1 or more eggs of helminth parasites. There was no significant difference in the distribution of eggs in the soil samples from the 3 areas sampled (c2=0.046, df=2, P>0.05). Ascaris species were the dominant parasite eggs found. Of the 180 faecal samples examined, 107 (59.4 %) were positive for 1 or more eggs of helminth parasites. Chi-square analysis showed no significant difference in the level of infection of different animal faeces sampled (c2=5.74, df=4, P>0.05). Ascaris species were again the dominating helminth parasite eggs found. There was also no significant difference in the prevalence of helminth eggs in the animal faecal samples from the 3 areas sampled (c2=5.99, df=4, P>0.05). A total of 1908 ectoparasites was recovered (ticks: 32.80 %; lice: 22.43 %; fleas: 22.06% and mite: 22.69 %). There was no significant difference in infestation animals between sexes (c2=0.10, df=4, P>0.05). The predominant genus encountered for ticks were Amblyomma, while Linognathus (43.90%), Ctenocephalides (97.38%) and Sarcoptes (58.89 %) were most predominant for lice, fleas and mites respectively. The public health implications of the findings, especially as these relate to the increasing incidence and prevalence of zoonotic infections, are discussed.


2011 ◽  
Vol 44 (4) ◽  
pp. 451-456 ◽  
Author(s):  
Charliana Aragão Damasceno Casal ◽  
Mayra Oliveira da Silva ◽  
Igor Brasil Costa ◽  
Eliete da Cunha Araújo ◽  
Tereza Cristina de Oliveira Corvelo

INTRODUCTION: Although control measures of maternal and congenital syphilis are available in Brazil, difficulties exist within the healthcare network in providing a laboratory diagnosis of the infection during the prenatal period. The objective of this study was to confirm the presence of Treponema pallidum by PCR in women with positive VDRL serology and lethal pregnancy outcomes, i.e., abortion, stillbirth and neonatal death. METHODS: A retrospective study was conducted on VDRLseroreactive women with lethal pregnancy outcomes admitted to the Fundação Santa Casa de Misericórdia do Pará (FSCM-PA) between January and July 2004. Serum samples and DNA from whole blood were obtained at the time of screening by the VDRL test. These samples were analyzed by IgG ELISA, IgM FTA-Abs and simple PCR (polA). RESULTS: During the study period, 0.7% (36/4,912) of women with lethal pregnancy outcomes presented a positive VDRL test. The polAgene was amplified in 72.7% (24/33) of these women, with 55.6% (20/36) and 94.4% (34/36) presenting IgM and IgG antibodies against T. pallidum, respectively. Comparison of these results showed a significant difference, with agreement between the PCR and IgM FTA-Abs results, suggesting that maternal syphilis was an active infection. No basic cause of death of the conceptus was reported in 97.2% (35/36) of cases. Among women who were submitted to the VDRL test during the prenatal period, only four of the nine seroreactive patients underwent treatment. CONCLUSIONS: The high frequency of syphilis in the group studied indicates the fragility of the service of infection diagnosis, treatment and monitoring, compromising epidemiological control.


Author(s):  
Enas Sh. Khater ◽  
Abd Alazim A. Al- Faki

Clostridium difficile infections (CDIs) is considered healthcare-associated infections which cause watery diarrhea to long stayed hospitalized patients and cause increased mortality rate. Aim: Detection of the prevalence and risk factors of C. difficile in Al Quwayiyah General hospital, Riyadh, Kingdom of Saudi Arabia and compairing between GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC, (QCC-EIA) in detection of C. difficile infection and toxicity Materials and Methods: A cross sectional and prospective study was performed for one year started from June 2019 to June 2020. The data collected include demographic, laboratory and clinical data. A total of 104 stool samples were collected from patients presented with diarrhea. GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC (QCC-EIA) were conducted to each stool sample. Results: Only 15(14.4%) of the 104 studied patients had CDI while 89 (85.6%) were non CDI patients, 13 (86.7%) of the CDI patients were males and 2 (13.3%) were females with mean age for CDI cases 61 (±19.9), while non CDI cases involved 55(61.8%) were males and 34 (38.2%) were females with mean age for cases of non CDI, 60 (±18.7) years. Of the CDI and non CDI cases respectively 12 (80%) and 14(15.7%) had fever, 5 (27%) and 6 (6.7%) had vomitting and 7 (46.7%) and 12 (13.5%) of cases had abdominal pain. There was statistical significant difference between patients with fever while no statistical significant difference regarding vomitting and abdominal pain. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics, while There was no statistical significant difference between cardiac disease, cerebrovascular disease, diabetes, pulmonary disease, hepatic disease and Renal disease. Gene expert PCR detected 15/104(14.4%) as positive CDI while QCC-EIA detected 21/104 (20.5%) as positive CDI. On comparison between gene expert PCR technique and QCC-EIA the sensitivity of QCC-EIA was 100%, while the specificity was 91%. The Positive Predictive Value was 74%, while the Negative Predictive Value was 100%. Conclusion: The C. difficile infection prevalence rate in the hospital was 14.4%. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics. The QCC-EIA can be used as a screening test for the detection of C. difficile toxin in stool samples but should be confirmed with a PCR assay or another confirmatory test Due to its decreased specificity.


2018 ◽  
Vol 56 (8) ◽  
Author(s):  
Alice Banz ◽  
Aude Lantz ◽  
Brigitte Riou ◽  
Agnès Foussadier ◽  
Mark Miller ◽  
...  

ABSTRACT Guidelines recommend the use of an algorithm for the laboratory diagnosis of Clostridium difficile infection (CDI). Enzyme immunoassays (EIAs) detecting C. difficile toxins cannot be used as standalone tests due to suboptimal sensitivity, and molecular tests suffer from nonspecificity by detecting colonization. Sensitive immunoassays have recently been developed to improve and simplify CDI diagnosis. Assays detecting CD toxins have been developed using single-molecule array (SIMOA) technology. SIMOA performance was assessed relative to a laboratory case definition of CDI defined by positive glutamate dehydrogenase (GDH) screen and cell cytotoxicity neutralizing assay (CCNA). Samples were tested with SIMOA assays and a commercial toxin EIA to compare performance, with discrepancy resolution using a commercial nucleic acid-based test and a second cell cytotoxicity assay. The SIMOA toxin A and toxin B assays showed limits of detection of 0.6 and 2.9 pg/ml, respectively, and intra-assay coefficients of variation of less than 10%. The optimal clinical thresholds for the toxin A and toxin B assays were determined to be 22.1 and 18.8 pg/ml, respectively, with resultant sensitivities of 84.8 and 95.5%. In contrast, a high-performing EIA toxin test had a sensitivity of 71.2%. Thus, the SIMOA assays detected toxins in 24% more samples with laboratory-defined CDI than the high performing toxin EIA (95% [63/66] versus 71% [47/66]). This study shows that SIMOA C. difficile toxin assays have a higher sensitivity than currently available toxin EIA and have the potential to improve CDI diagnosis.


2018 ◽  
Vol 39 (4) ◽  
pp. 434-438 ◽  
Author(s):  
Donna M. Schora ◽  
Lance R. Peterson ◽  
Elena A. Usacheva

OBJECTIVEThe impact of storage on stability and detection ofClostridium difficiletoxins in feces is poorly understood. The objective of this study was to investigate the immunological stability ofC. difficiletoxins in clinical stool specimens under different storage conditions by evaluating this stability using toxin detection by enzyme immunoassay (EIA).METHODSStool specimens positive forC. difficileinfection (CDI) by quantitative polymerase chain reaction (qPCR) were used for EIA testing with theC. difficileTox A/B II kit. The EIA-positive specimens were stored aerobically under refrigerated (4–10°C) and frozen (−30°C and −80°C) conditions. Measurement of toxin quantity was conducting using optical density (OD) on days 0, 14, 30, 60, 90, and 120 of storage.RESULTSClostridium difficiletoxins demonstrated good detection in undiluted stool specimens by EIA up to 120 days of storage. Good detection of the toxins was observed in diluted samples at refrigerated and −80°C temperatures. Dilution detrimentally affected toxin detection at −30°C.CONCLUSIONStorage of undiluted clinical stool specimens at refrigerated, −30°C, and −80°C temperatures for up to 120 days has no discernible effect on the immunological stability ofC. difficilecytotoxins. However, storage at −30°C has a detrimental effect onC. difficiletoxin stability in diluted specimens.Infect Control Hosp Epidemiol2018;39:434–438


Gut ◽  
2018 ◽  
Vol 67 (12) ◽  
pp. 2107-2115 ◽  
Author(s):  
Sofie Ingdam Halkjær ◽  
Alice Højer Christensen ◽  
Bobby Zhao Sheng Lo ◽  
Patrick Denis Browne ◽  
Stig Günther ◽  
...  

ObjectiveIBS is associated with an intestinal dysbiosis and faecal microbiota transplantation (FMT) has been hypothesised to have a positive effect in patients with IBS. We performed a randomised, double-blind placebo-controlled trial to investigate if FMT resulted in an altered gut microbiota and improvement in clinical outcome in patients with IBS.DesignWe performed this study in 52 adult patients with moderate-to-severe IBS. At the screening visit, clinical history and symptoms were assessed and faecal samples were collected. Patients were randomised to FMT or placebo capsules for 12 days and followed for 6 months. Study visits were performed at baseline, 1, 3 and 6 months, where patients were asked to register their symptoms using the IBS-severity scoring system (IBS-SSS) and IBS-specific quality of life (IBS-QoL). Prior to each visit, faecal samples were collected.ResultsA significant difference in improvement in IBS-SSS score was observed 3 months after treatment (p=0.012) favouring placebo. This was similar for IBS-QoL data after 3 months (p=0.003) favouring placebo. Patients receiving FMT capsules had an increase in faecal microbial biodiversity while placebos did not.ConclusionIn this randomised double-blinded placebo-controlled study, we found that FMT changed gut microbiota in patients with IBS. But patients in the placebo group experienced greater symptom relief compared with the FMT group after 3 months. Altering the gut microbiota is not enough to obtain clinical improvement in IBS. However, different study designs and larger studies are required to examine the role of FMT in IBS.Trial registration numberNCT02788071.


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