scholarly journals Evaluation of StrepBSelectChromogenic Medium and the Fast-Track Diagnostics Group B Streptococcus (GBS) Real-Time PCR Assay Compared to Routine Culture for Detection of GBS during Antepartum Screening

2017 ◽  
Vol 55 (7) ◽  
pp. 2137-2142 ◽  
Author(s):  
Deirdre L. Church ◽  
Heather Baxter ◽  
Tracie Lloyd ◽  
Oscar Larios ◽  
Daniel B. Gregson
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fast Track ◽  
Group B Streptococcus ◽  
Culture Method ◽  
Predictive Values ◽  
Content Type ◽  
Standard Culture ◽  
Group B ◽  
The Cost

ABSTRACTLife-threatening infection in neonates due to group BStreptococcus(GBS) is preventable by screening of near-term pregnant women and treatment at delivery. A total of 295 vaginal-rectal swabs were collected from women attending antepartum clinics in Calgary, Alberta, Canada. GBS colonization was detected by the standard culture method (Strep B Carrot Broth subcultured to blood agar with a neomycin disk) and compared to recovery with Strep Group B Broth (Dalynn Biologicals) subcultured to StrepBSelectchromogenic medium (CM; Bio-Rad Laboratories) and the Fast-Track Diagnostics GBS real-time PCR (quantitative PCR [qPCR]) assay (Phoenix Airmid Biomedical Corp.) performed with broth-enriched samples and the Abbottm2000sp/m2000rt system. A total of 62/295 (21%) women were colonized with GBS; 58 (19.7%) cases were detected by standard culture, while CM and qPCR each found 61 (20.7%) cases. The qPCR and CM were similar in performance, with sensitivities, specificities, and positive and negative predictive values of 98.4 and 98.4%, 99.6 and 99.6%, 98.4 and 98.4%, and 99.6 and 99.6%, respectively, compared to routine culture. Both qPCR and CM would allow more rapid reporting of routine GBS screening results than standard culture. Although the cost per test was similar for standard culture and CM, the routine use of qPCR would cost approximately four times as much as culture-based detection. Laboratories worldwide should consider implementing one of the newer methods for primary GBS testing, depending on the cost limitations of different health care jurisdictions.

Evaluation of Culture Method and Real-Time PCR for Detection of Vaginal Group B Streptococcus Colonization in Pregnant at the Last Trimester

2014 ◽  
Vol 56 (2) ◽  
pp. 80
Author(s):  
Samer ALWEDYAN ◽  
Ramazan GMRAL ◽  
mit GOKTOLGA ◽  
Fatih SAHINER ◽  
Orhan BEDIR ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Group B Streptococcus ◽  
Culture Method ◽  
Group B

scholarly journals Development of Conventional and Real-Time PCR Assays for the Rapid Detection of Group B Streptococci

2000 ◽  
Vol 46 (3) ◽  
pp. 324-331 ◽  
Author(s):  
Danbing Ke ◽  
Christian Ménard ◽  
François J Picard ◽  
Maurice Boissinot ◽  
Marc Ouellette ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Culture Method ◽  
Conventional Pcr ◽  
Pcr Assay ◽  
Group B Streptococci ◽  
Standard Culture ◽  
Pcr Assays ◽  
Group B

Abstract Background: Group B streptococci (GBS), or Streptococcus agalactiae, are the leading bacterial cause of meningitis and bacterial sepsis in newborns. Currently available rapid methods to detect GBS from clinical specimens are unsuitable for replacement of culture methods, mainly because of their lack of sensitivity. Methods: We have developed a PCR-based assay for the rapid detection of GBS. The cfb gene encoding the Christie-Atkins-Munch-Petersen (CAMP) factor was selected as the genetic target for the assay. The PCR primers were initially tested by a conventional PCR method followed by gel electrophoresis. The assay was then adapted for use with the LightCyclerTM. For this purpose, two fluorogenic adjacent hybridization probes complementary to the GBS-specific amplicon were designed and tested. In addition, a rapid sample-processing protocol was evaluated by colony-forming unit counting and PCR. A total of 15 vaginal samples were tested by both standard culture method and the two PCR assays. Results: The conventional PCR assay was specific because it amplified only GBS DNA among 125 bacterial and fungal species tested, and was able to detect all 162 GBS isolates from various geographical areas. This PCR assay allowed detection of as few as one genome copy of GBS. The real-time PCR assay was comparable to conventional PCR assay in terms of sensitivity and specificity, but it was more rapid, requiring only ∼30 min for amplification and computer-based data analysis. The presence of vaginal specimens had no detrimental effect on the sensitivity of the PCR with the sample preparation protocol used. All four GBS-positive samples identified by the standard culture method were detected by the two PCR assays. Conclusion: These assays provide promising tools for the rapid detection and identification of GBS.

scholarly journals Real-Time PCR Assay Provides Reliable Assessment of Intrapartum Carriage of Group B Streptococcus

2010 ◽  
Vol 48 (9) ◽  
pp. 3095-3099 ◽  
Author(s):  
M. J. Alfa ◽  
S. Sepehri ◽  
P. De Gagne ◽  
M. Helawa ◽  
G. Sandhu ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Group B Streptococcus ◽  
Pcr Assay ◽  
Reliable Assessment ◽  
Group B

scholarly journals Combined Loop-Mediated Isothermal Amplification Assays for Rapid Detection and One-Step Differentiation of Campylobacter jejuni and Campylobacter coli in Meat Products

2021 ◽  
Vol 12 ◽  
Author(s):  
Antonia Kreitlow ◽  
André Becker ◽  
Marwa F. E. Ahmed ◽  
Sophie Kittler ◽  
Ulrich Schotte ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Lamp Assay ◽  
Culture Method ◽  
Campylobacter Coli ◽  
Standard Culture ◽  
One Step ◽  
Meat Samples ◽  
Minced Meat

A loop-mediated isothermal amplification (LAMP) assay system was established, allowing rplD gene-based simultaneous detection of Campylobacter jejuni and Campylobacter coli in enriched meat products. Additionally, one-step differentiation of target species on agar plates was enabled by cdtC gene- and gyrA gene-based duplex LAMP. Both the rplD and cdtC–gyrA LAMP assays amplified the target sequences in all 62 C. jejuni and 27 C. coli strains used for determining inclusivity and revealed 100% exclusivity toward 85 tested non-target species. Throughout the entire experiments, C. jejuni and C. coli strains were 100% distinguishable by melting curves of cdtC and gyrA LAMP products. After 24-h enrichment, the rplD LAMP assay reliably detected initial inoculation levels of 10–100 CFU/g in artificially contaminated minced meat. Investigation of naturally contaminated meat samples revealed a diagnostic accuracy of 95% toward real-time PCR and 94.1% toward the standard culture method applying the 24-h incubation period. Diagnostic sensitivity and specificity, and positive and negative predictive values were 89.8, 100, 100, and 91.2%, respectively, when measured against real-time PCR, and 89.6, 98.1, 97.7, and 91.2%, respectively, when measured against the standard culture method. After 48-h enrichment, the detection limit of the rplD LAMP assay improved to initial inoculation levels of 1–10 CFU/g in artificially contaminated minced meat. Applying the 48-h incubation period on naturally contaminated meat samples resulted in 100% concordant results between rplD LAMP, real-time PCR, and the standard culture method. The established LAMP assay system was proved to be suitable for rapid meat sample screening. Furthermore, it constitutes a promising tool for investigating other Campylobacter sources and could therefore make a valuable contribution to protect consumers from foodborne illness.

Comparison of group B strep IDI-StrepB real-time PCR assay to standard culture

2005 ◽  
Vol 193 (6) ◽  
pp. S186
Author(s):  
Kristin Atkins (F) ◽  
Robyn Atkinson ◽  
Anthony Shanks ◽  
W. Michael Dunne ◽  
Gilad Gross
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Assay ◽  
Standard Culture ◽  
Group B

scholarly journals Evaluation of the ELITe InGenius PCR Platform for Detection of Mycoplasma pneumoniae

2019 ◽  
Vol 57 (6) ◽  
Author(s):  
Arthur H. Totten ◽  
Sixto M. Leal ◽  
Amy E. Ratliff ◽  
Li Xiao ◽  
Donna M. Crabb ◽  
...  
Keyword(s):  
Real Time ◽  
Mycoplasma Pneumoniae ◽  
Real Time Pcr ◽  
Growth Media ◽  
Test Accuracy ◽  
Analytical Sensitivity ◽  
Pcr Assay ◽  
Predictive Values ◽  
Content Type ◽  
Significant Difference

ABSTRACT Mycoplasma pneumoniae is the leading cause of bacterial community-acquired pneumonia in persons of all ages. Due to the fastidious nature of this bacterium and the necessary specialized growth media, nucleic acid amplification testing is currently the most reliable means for patient diagnostics. Analytical sensitivity, specificity, reproducibility, and clinical performance of the ELITe InGenius automated PCR platform with its MGB Alert M. pneumoniae real-time PCR research use only reagents (ELITechGroup, Inc., Bothell, WA) were compared with those of a laboratory-developed real-time PCR assay targeting repMp1 for detection of M. pneumoniae. The ELITe InGenius PCR assay successfully detected 31 distinct M. pneumoniae clinical isolates and reference strains, and there was no cross-reactivity with other mollicutes, Gram-positive bacteria, or Gram-negative bacteria. In testing 223 clinical samples, the ELITe InGenius PCR showed 95.79% and 99.22% positive and negative agreement with the repMp1 assay, respectively. Additionally, the ELITech platform showed 98.91% positive and 96.95% negative predictive values, and there was no significant difference detected between the two assays (McNemar’s test, P = 0.375). The ELITe InGenius PCR assay limit of detection was 0.16 CFU/PCR test or 4.16 genome copies (GCs)/test. Accuracy, instrument ease-of-use, and decreased hands-on time make the ELITe InGenius platform suitable for detection of M. pneumoniae directly from clinical specimens.

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