Culture-based method with performance comparable to that of PCR-based methods for detection of group B Streptococcus in screening samples from pregnant women.
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ABSTRACT:
SUBMITTER: Berg BR
PROVIDER: S-EPMC3666823 | biostudies-other | 2013 Apr
REPOSITORIES: biostudies-other
Publications
Culture-based method with performance comparable to that of PCR-based methods for detection of group B Streptococcus in screening samples from pregnant women.
Berg Benjamin R BR Houseman Jeana L JL Garrasi Michelle A MA Young Carol L CL Newton Duane W DW
Journal of clinical microbiology 20130123 4
We compared five approaches for group B streptococcus (GBS) detection: three culture-based methods and two methods using broth-enhanced real-time PCR. Carrot broth-enhanced subculture to GBS Detect (Hardy Diagnostics, Santa Maria, CA) exhibited sensitivity and specificity comparable to carrot broth- and LIM broth-enhanced real-time PCRs. ...[more]
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Project description:BACKGROUND: Group B streptococcus (GBS) is one of the most important cause of morbidity and mortality among newborns especially in developing countries. It has been shown that the screening approach rather than the identification of maternal clinical risk factors for early-onset neonatal GBS disease is more effective in preventing early-onset GBS neonatal disease. The objective of this study was to detect GBS among clinical samples of women using PCR and standard microbiological culture. METHODS: Samples were taken from 375 women at 28-38 weeks of gestation during six month from January 15 till June 15, 2011 from a hospital in Tehran, Iran. Samples were tested by standard culture using Todd-Hewitt broth, blood agar and by PCR targeting the cfb gene. RESULTS: Among the 375 women, 35 (9.3%) were identified as carriers of group B streptococci on the basis of the results of the cultures of specimens, compared to 42 (11.2 %) on the basis of PCR assay. CONCLUSION: We found that GBS can be detected rapidly and reliably by a PCR assay in vaginal secretions from women at the time of delivery. This study also showed that the rate of incidence of GBS is high in Iranian women.
| S-EPMC3481701 | biostudies-literature
Project description:Group B streptococcus (GBS) is a major cause of serious infections in neonates. The 2002 revised guidelines of the Centers for Disease Control and Prevention (CDC) for the prevention of perinatal GBS disease recommend that all pregnant women be screened for GBS carriage at between 35 and 37 weeks of gestation and that intrapartum antibiotic prophylaxis be given to carriers. We studied the performances of four different GBS detection assays in the context of antenatal screening. Between May and August 2004, the 605 vaginorectal swab specimens received at our bacteriology laboratory for GBS antenatal detection were tested by the four assays. The standard culture method was done according to the CDC recommendations. The three experimental assays performed with the growth from the selective enrichment (LIM) broth (Todd-Hewitt broth with 15 mug/ml nalidixic acid and 10 mug/ml colistin) after overnight incubation were a GBS antigen detection assay (PathoDx) and two PCR assays (for cfb and scpB). The most accurate assay was the scpB PCR (sensitivity, 99.6%; specificity, 100%), followed by the cfb PCR (sensitivity, 75.3%; specificity, 100%), GBS antigen detection (sensitivity, 57.3%; specificity, 99.5%), and standard culture (sensitivity, 42.3%; specificity, 100%). The GBS antigen detection assay was found to be more sensitive than the standard culture method, and moreover, the assay has a low cost and is easy to perform in all obstetrical centers which have access to the most basic of diagnostic microbiology services. We believe that antigen detection on incubated LIM broth should replace the standard culture method for screening for GBS carriage at 35 to 37 weeks of gestation. The impact of the greater sensitivities of PCR assays on the diminution of neonatal GBS infections remains to be demonstrated.
| S-EPMC1393163 | biostudies-literature
Project description:BackgroundStreptococcus agalactiae (group B streptococcus; GBS) is a significant cause of perinatal and neonatal infections worldwide. To detect GBS colonization in pregnant women, the CDC recommends isolation of the bacterium from vaginal and anorectal swab samples by growth in a selective enrichment medium, such as Lim broth (Todd-Hewitt broth supplemented with selective antibiotics), followed by subculture on sheep blood agar. However, this procedure may require 48 h to complete. We compared different sampling and culture techniques for the detection of GBS.MethodsA total of 300 swabs was taken from 100 pregnant women at 35-37 weeks of gestation. For each subject, one rectovaginal, one vaginal and one rectal ESwab were collected. Plating onto Columbia CNA agar (CNA), group B streptococcus differential agar (GBSDA) (Granada Medium) and chromID Strepto B agar (CA), with and without Lim broth enrichment, were compared. The isolates were confirmed as S. agalactiae using the CAMP test on blood agar and by molecular identification with tDNA-PCR or by 16S rRNA gene sequence determination.ResultsThe overall GBS colonization rate was 22%. GBS positivity for rectovaginal sampling (100%) was significantly higher than detection on the basis of vaginal sampling (50%), but not significantly higher than for rectal sampling (82%). Direct plating of the rectovaginal swab on CNA, GBSDA and CA resulted in detection of 59, 91 and 95% of the carriers, respectively, whereas subculturing of Lim broth yielded 77, 95 and 100% positivity, respectively. Lim broth enrichment enabled the detection of only one additional GBS positive subject. There was no significant difference between GBSDA and CA, whereas both were more sensitive than CNA. Direct culture onto GBSDA or CA (91 and 95%) detected more carriers than Lim broth enrichment and subculture onto CNA (77%). One false negative isolate was observed on GBSDA, and three false positives on CA.ConclusionsIn conclusion, rectovaginal sampling increased the number GBS positive women detected, compared to vaginal and/or rectal sampling. Direct plating on CA and/or GBSDA provided rapid detection of GBS that was at least as sensitive and specific as the CDC recommended method of Lim broth subcultured onto non chromogenic agar.
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Project description:BACKGROUND:Streptococcus Group B (GBS) colonization in pregnant women is the most important risk factor for newborn disease due to vertical transmission during delivery. GBS colonization during pregnancy has been implicated as a leading cause of perinatal infections. Traditionally, pregnant women are screened for GBS between 35 and 37 weeks of gestation. However, antenatal culture-based screening yields no information on GBS colonization status and offers low predictive value for GBS colonization at delivery. Numerous assays have been evaluated for GBS screening in an attempt to validate a fast and efficient method. The aim of this study was to compare bacteria isolation by culture and two qPCR techniques, targeting sip and cfb genes, respectively, for detecting colonizing GBS. METHODS:Cultures - the gold-standard technique, a previous qPCR technique targeting the sip gene, and a new proposed qPCR assay targeting the cfb gene were evaluated as diagnostic tools on 320 samples. RESULTS:Considering cultures as the gold standard, the evaluated qPCR method detected 75 out of 78 samples, representing a sensitivity of 93.58% (95% confidence interval (CI), 90.89-96.27) and specificity of 94.62% (95% CI, 91.78-97.46). However, an additional analysis was performed for true positives that included not only samples showing positives by culture but samples showing positive for both qPCR assays. The sensitivity and specificity were recalculated including these discrepant samples and a total of 89 samples were considered as positive, giving a prevalence of 27.81%. With this new analysis, the qPCR targeting the cfb gene showed a sensitivity of 95.5% (95% CI, 88.65-98.59) and specificity of 99.13% (95% CI, 96.69-99.97). CONCLUSIONS:The new qPCR method is a sensitive and specific assay for detecting GBS colonization and represents a valuable tool for identifying candidates for intrapartum antibiotic prophylaxis. Cultures should be retained as the reference and the routine technique because of its specificity and cost analysis ratio, but it would be convenient to introduce PCR techniques to check negative culture samples or when an urgent detection is required to reduce risk of infection among infants.
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Project description:BackgroundSeveral in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans.MethodsA total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae.ResultsAll used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains.ConclusionsThe in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.
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Project description:Group B streptococcus (GBS), which commonly colonizes the female genital tract and rectum, can cause infections in newborns with varying severity, possibly leading to death. The aim of the present study was to evaluate Hitchens-Pike-Todd-Hewitt (HPTH) medium performance for GBS screening in pregnant women. A descriptive analytical cross-sectional study was performed with 556 pregnant women, of which 496 were at 35-37 weeks of gestation and 60 were at ? 38 weeks of gestation. The study was conducted from September 2011 to March 2014 in northern Paraná, Brazil. Vaginal and anorectal clinical specimens from each pregnant woman were plated on sheep blood agar (SBA) and seeded on HPTH medium and Todd-Hewitt enrichment broth. Of the 496 pregnant women at 35-37 weeks of gestation, 141 (28.4%) were positive for GBS, based on the combination of the three culture media and clinical specimens. The GBS colonization rates that were detected by each medium were 22.2% for HPTH medium, 21.2% for SBA, and 13.1% for Todd-Hewitt enrichment broth. Of the 60 pregnant women at ? 38 weeks of gestation, seven (11.7%) were positive for GBS. These results demonstrate that HPTH medium and SBA were more sensitive than Todd-Hewitt enrichment broth for GBS screening in pregnant women and good GBS recovery in culture, indicating that the two media should be used together for vaginal and anorectal specimens.
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Project description:The objective of this study was to estimate the sensitivity and specificity of a culture method and a polymerase chain reaction (PCR) method for detection of two Campylobacter species: C. jejuni and C. coli. Data were collected during a 3-year survey of UK broiler flocks, and consisted of parallel sampling of caeca from 436 batches of birds by both PCR and culture. Batches were stratified by season (summer/non-summer) and whether they were the first depopulation of the flock, resulting in four sub-populations. A Bayesian approach in the absence of a gold standard was adopted, and the sensitivity and specificity of the PCR and culture for each Campylobacter subtype was estimated, along with the true C. jejuni and C. coli prevalence in each sub-population. Results indicated that the sensitivity of the culture method was higher than that of PCR in detecting both species when the samples were derived from populations infected with at most one species of Campylobacter. However, from a mixed population, the sensitivity of culture for detecting both C. jejuni or C. coli is reduced while PCR is potentially able to detect both species, although the total probability of correctly identifying at least one species by PCR is similar to that of the culture method.
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Project description:ObjectiveTo determine the validity of a novel Group B Streptococcus (GBS) diagnostic assay for the detection of GBS in antepartum patients.Study designWomen were screened for GBS colonization at 35 to 37 weeks of gestation. Three vaginal-rectal swabs were collected per patient; two were processed by traditional culture (commercial laboratory versus in-house culture), and the third was processed by an immunoblot-based test, in which a sample is placed over an antibody-coated nitrocellulose membrane, and after a six-hour culture, bound GBS is detected with a secondary antibody.Results356 patients were evaluated. Commercial processing revealed a GBS prevalence rate of 85/356 (23.6%). In-house culture provided a prevalence rate of 105/356 (29.5%). When the accelerated GBS test result was compared to the in-house GBS culture, it demonstrated a sensitivity of 97.1% and a specificity of 88.4%. Interobserver reliability for the novel GBS test was 88.2%.ConclusionsThe accelerated GBS test provides a high level of validity for the detection of GBS colonization in antepartum patients within 6.5 hours and demonstrates a substantial agreement between observers.
| S-EPMC3590750 | biostudies-literature
Project description:The use of some nucleic acid amplification tests (NAATs) for the diagnosis of group A Streptococcus (GAS) pharyngitis allows laboratories to adopt single-tiered testing without reflex culture. However, centralization may delay the delivery of actionable information to the bedside, particularly in the outpatient setting. We describe two novel workflows at our institution and their effect on in-lab turnaround time (TAT) at a tertiary care microbiology lab. Laboratory records were extracted, and relevant data were analyzed after the implementation of qualitative in vitro diagnostic testing for GAS with the Xpert Xpress Strep A assay, performed using the GeneXpert Infinity-48s. Workflow optimization steps studied included: (i) direct specimen submission to the microbiology laboratory via the pneumatic tube system and (ii) autoverification of GAS NAAT results in the laboratory information system. Between April 2018 and October 2018, 2,595 unique specimens were tested for GAS by PCR. Of these, 2,523 were included in the final analysis. Linear regression established that the total in-lab TAT was significantly reduced by direct specimen submission to the microbiology laboratory, autoverification, and processing during the night shift. We describe two workflow optimization methods that reduced the in-lab TAT for GAS NAAT. Although microbiology labs historically use manual processes, the advent of total laboratory automation and the adoption of on-demand NAATs will allow for more streamlined processing of microbiology specimens. It may be beneficial to consider instrument interfacing and specimen processing optimization during the early phases of implementation planning for NAATs in the microbiology laboratory.
| S-EPMC6711925 | biostudies-literature
Project description:AIMS:Group B streptococcus (GBS) is one of the most frequent bacterial pathogens causing invasive infections in neonates. It can be transmitted from colonised mother to neonates around delivery. Screening strategies for GBS during pregnancy include either universal culture-based or risk-based screening. The present study aimed to assess the knowledge, attitude and current practices of pregnant women towards GBS screening in Al-Madinah City, Saudi Arabia. METHODS:A hospital-based cross-sectional study was conducted at Madinah Maternity and Children Hospital, Al-Madinah, Saudi Arabia, from May to July 2018. Participants were recruited from postnatal wards. Participants were interviewed using a previously published validated survey that was divided into the following domains: demographic data, knowledge, experience and attitude towards different GBS screening strategies. RESULTS:A total of 377 out of 450 women (response rate 83.7%) were enrolled. The results showed that the overall mean knowledge score of the pregnant women towards GBS screening was 59.8%. Majority of the women (66.8%) were not aware of the GBS bacterium, while 86.5% of them had never been informed of GBS risk assessment during their pregnancies. The mean knowledge score among women who were aware of GBS (62.8%) was significantly higher than that among women who were not (58.4%) (p=0.015). However, majority of the pregnant women (61.8%) showed preference for universal culture-based GBS screening strategy over risk-based strategy. CONCLUSION:The study results have concluded that the level of awareness and knowledge about GBS among pregnant women were relatively poor; however, majority of the pregnant women prefer universal culture-based screening.
| S-EPMC7045259 | biostudies-literature