Evaluation of Trans-Vag Broth,Colistin-Nalidixic Agar,and CHROMagar StrepB for Detection of Group B Streptococcus in Vaginal and Rectal Swabs from Pregnant Women in South Africa

Maternal vaginal colonization with group B streptococcus (GBS) is a major risk factor for invasive GBS infection in newborns. The CDC-recommended method for detecting GBS colonization is to culture vaginal and rectal swabs in a selective broth followed by subculture on blood agar or a selective medium. A high incidence of antimicrobial resistance in the fecal microflora can compromise the recovery of GBS from the selective broth. Here, we compared CHROMagar StrepB (CA), Columbia colistin-nalidixic agar (CNA), and Trans-Vag selective broth enrichment for the isolation of GBS from 130 vaginal and 130 rectal swabs from pregnant women. The swabs were randomized for plating first on either CA or CNA, and they then were inoculated in Trans-Vag broth. GBS was cultured from 37.7% of the vaginal swabs and 33.1% of the rectal swabs. There were no differences in the detection rates for the vaginal swabs between CA (31.5%), CNA (26.2%), and the selective broth (30.0%). The sensitivities in relation to a composite score were 83.7%, 69.4%, and 79.6%, respectively. However, recovery of GBS from the rectal swabs was significantly higher from CA (29.2%; P < 0.0001) and CNA (23.8%; P = 0.002) than from the selective broth (9.2%). The sensitivities were 88.4%, 72.1%, and 27.9%, respectively. The order of plating on the solid medium was significant (P = 0.003), with GBS detection rates of 30.8% and 24.6% when swabs were plated first and second, respectively. These findings show that a selective broth is not suitable for the recovery of GBS from rectal swabs in settings such as ours, due to masking of the GBS colonies by persistent microflora.     

Comparison of two culture media and three sampling techniques for sensitive and rapid screening of vaginal colonization by group B streptococcus in pregnant women

The Centers for Disease Control and Prevention (CDC) recommend universal screening of all pregnant women between 35 and 37 weeks of gestation for group B streptococci (GBS) by use of a selective broth medium. Recent reports suggest that Granada medium can be used for rapid and direct visual identification of GBS colonies. However, studies comparing the Granada medium method to the selective broth method are few, and while some report comparable sensitivities, others have found significant differences in detection rates between the two methods. This prospective study compared a method using Granada agar to a Todd-Hewitt broth method with subculture to blood agar in order to determine which GBS detection method is more sensitive and less labor-intensive and has a more rapid turnaround time. Detection rates for three sampling techniques (rectovaginal, vaginal only, and cervical only) were also compared. Consecutive specimens for GBS screening received over a 6-month period from 1,635 pregnant women were included. Overall, GBS was detected in 390 (23.8%) women. The Granada medium gave positive results for 348 of these women, and the selective broth gave positive results for 385, indicating sensitivities of 89.2% for the Granada medium and 98.7% for the selective broth. These findings show that the Granada medium method is less sensitive than the selective broth method and should not replace it as the only method for screening pregnant women for GBS. However, the Granada medium method reduced detection time to 1 day and also reduced the use of ancillary tests in approximately 90% of positive cases. Additionally, no significant differences were noted in the detection rates with rectovaginal, vaginal, and cervical specimens.     

Evaluation of Group B Streptococcus Differential Agar for detection and isolation of Streptococcus agalactiae

In total, 320 vaginal or rectal swabs were cultured on Granada medium (GM) or Group B Streptococcus Differential Agar (GBSDA), and were also inoculated into LIM broth (Todd-Hewitt broth supplemented with selective antibiotics), for detection of group B Streptococcus (GBS). Overall, GBS isolates were detected on 53 of the 320 swabs; 47 of these isolates grew on both GM and GBSDA, five only on GBSDA, and one only following subculture from LIM broth. GBSDA appears to be a valid alternative to GM for the growth of GBS isolates from pregnant women.     

Group B streptococcus prevalence in pregnant women from North-Eastern Italy: advantages of a screening strategy based on direct plating plus broth enrichment

AIMS: To assess the sensitivity of a combined selective broth enrichment technique plus selective plating for the detection of group B streptococcus (GBS) colonisation in a large cohort of pregnant women from North-Eastern Italy. METHODS: During 2002-2005, 5020 pregnant women were screened between the 35th and the 37th week of gestation. A lower vaginal sample and a rectal sample were collected and inoculated onto LIM broth and a selective colistin aztreonam blood agar plate (CAP). Direct agar plates were examined after 18-24 hours and, if negative, after 48 hours. LIM broth was subcultured after 18-24 hours onto a Columbia blood agar plate. All colonies suggestive for GBS were submitted to phenotypic identification. RESULTS: 901 Women (17.9%) were positive for GBS. On 728 positive samples, corresponding to patients enrolled between 2003 and 2005, the results of selective direct plating and selective broth enrichment were compared. A total of 561 (77.1% of positive samples, corresponding to 13.9% of patients) were positive on direct selective agar; an additional 167 isolates (22.9% of samples, 4.1% of patients) were recovered from the LIM broth subculture. CONCLUSIONS: The prevalence of GBS carriage in this population-based study is a reliable estimate considering the sensitivity of the microbiological methods used, the rate of attendance of pregnant women to clinical and laboratory settings and the compliance to the protocol. Results confirm that the combination of selective enrichment broth and selective direct plating is a time-saving and sensitive method.     

Optimisation of Prenatal Group B Streptococcal Screening

The purpose of the study presented here was to confirm the high yield of group B streptococci (GBS) on Granada medium for the detection of pregnant GBS carriers and to compare the results with those obtained using standard Columbia blood agar at two participating centers in Belgium. Culture results of the vaginorectal swabs obtained at the two centers were also compared. A total of 1,142 samples (838 in Leuven and 304 in Bonheiden) obtained from consecutive pregnant women were cultured onto both media. Of all GBS carriers 84.7% were detected on Columbia blood agar and 93.4% on Granada agar (P<0.01, McNemar test). The addition of Granada agar was responsible for a 15% higher rate of detection of GBS carriers. As a result of this study, both participating hospitals will use a combination of Granada agar with Columbia blood agar for optimal GBS screening in the future.     

Use of GBS Media for Rapid Detection of Group B Streptococci in Vaginal and Rectal Swabs from Women in Labor

 In order to evaluate the differences in efficacy, three methods were used to detect group B streptococci (GBS) in women in labor. The recommended method for detecting GBS carriage in pregnant women is to culture vaginal and anorectal swabs in a selective broth medium and to subculture them onto blood agar. This method was compared with the use of GBS agar and GBS broth, both of which produce an orange pigment in response to GBS strains. A total of 319 women in labor were screened. Among the 638 specimens tested, 134 (21%) were positive in the selective Todd-Hewitt broth subcultured onto sheep blood agar, 133 (20.8%) were positive on the GBS agar and 126 (19.7%) were positive in the GBS broth. Altogether, 89 (27.9%) women in labor were found to be colonized with GBS; 87 (97.8%) of them were identified as carriers using the Todd-Hewitt broth, 87 (97.8%) with the GBS agar and 86 (96.6%) with the GBS broth. These results indicate that both GBS agar and GBS broth are reliable methods that can be used to screen for maternal and neonatal GBS colonization.     

Evaluation of Granada agar plate for detection of Streptococcus agalactiae in urine specimens from pregnant women

The Granada agar plate (GAP; Biomedics SL, Madrid, Spain) was evaluated for the detection of group B streptococci (GBS) in urine specimens from pregnant women submitted for testing for asymptomatic bacteriuria and was compared with blood agar (BA [Columbia agar with 5% sheep blood]; bioMérieux, Marcy l'Etoile, France). The GAP detected 103 out of 105 GBS, whereas BA detected only 50. Use of the GAP could be a good method for the detection of GBS in urine specimens from pregnant women.     

Evaluation of the Granada medium used for the determination of Streptococcus agalactiae at the eighth month of pregnancy

Granada medium (GM) was evaluated for the detection of group B streptococci (GBS) in vaginal swabs compared with the standard culture on selective blood agar (BA) and classical identification methods. From May to November 2002, samples from 325 pregnant women (34 to 37 weeks of gestation) were processed and 44 of these women (13.5%) carried GBS. Comparatively, GM was found more sensitive than the selective BA (95% versus 91%) in GBS recovery. The characteristic red-orange colonies produced by GBS are so specific that further identification is unnecessary. The technique is simple and results are available after overnight incubation, improving the time to reporting a GBS-positive result by at least 24 h. The inconvenience of anaerobic incubation of GM plates can be avoided when a cover slide is placed upon the inoculum because the same pigmentation is obtained under these aerobic conditions. This study confirms that the routine use of GM appears to be an accurate, easy and highly sensitive method of identification of GBS in pregnant women.     

Colonization rates and serotypes of group B streptococci isolated from pregnant women in a Korean tertiary hospital

In a study designed to provide data on the rates of maternal carriage of group B streptococci (GBS) in Korean women, vaginal, anorectal, and urethral swab specimens from 459 pregnant women and ear canal and umbilicus swabs from their 288 neonates were cultured with new Granada medium and selective Todd-Hewitt broth. Additionally, the serotypes of 64 isolates of GBS and the minimal inhibitory concentrations of seven antimicrobial agents for these isolates were determined. The rate of colonization by GBS in pregnant women and in their babies was 5.9% (27/459) and 0.7% (2/288), respectively. The rates of resistance of GBS isolated from pregnant women were 13.3% to clindamycin, 5% to erythromycin, and 98.3% to tetracycline. The majority of GBS isolates from pregnant women belonged to serotypes Ib (48.3%), la (24.1 %), and III (20.7%).     

Evaluation of StrepB carrot broth versus Lim broth for detection of group B Streptococcus colonization status of near-term pregnant women

The performance of StrepB Carrot Broth (SCB) versus group B Lim broth (LIM) for detection of group B streptococcus (GBS) colonization status in near-term pregnant women (35 to 37 weeks of gestation) was evaluated. Dually collected vaginal/rectal swabs from 279 women enrolled from a single large maternity clinic were analyzed. Fifty (18%) women were colonized by GBS according to both methods. SCB had excellent diagnostic performance compared to LIM, with sensitivity, specificity, positive predictive value, and negative predictive value of 92%, 100%, 100%, and 98.3%, respectively. Improved diagnostic efficiency due to direct reporting of GBS cases based on an orange color change in the SCB decreased overall labor and material costs.     

Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women

Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods for detecting GBS colonization.For a total of 100 pregnant women at 35-37 weeks of gestation, one rectovaginal ESwab each was collected. Eswab medium was inoculated into Lim broth, incubated for 24 h and plated onto chromID™ Strepto B agar (ChromAgar). DNA was extracted with the bioMérieux easyMAG platform, either directly from the rectovaginal ESwab or from Lim broth enrichment culture. Two different qPCR formats were compared, i.e. the hydrolysis probe format (Taqman, Roche) targeting the sip gene and the hybridization probe format (Hybprobe, Roche) targeting the cfb gene.Both qPCR techniques identified 33% of the women as GBS-positive. Only one culture-positive sample was qPCR-negative. QPCR directly on the sample significantly increased the number of women found to be GBS-positive (27%) compared to culture (22%). Moreover, the sensitivity of qPCR after Lim broth enrichment (33%) was again significantly higher than qPCR after DNA extraction directly from the rectovaginal swabs (27%).In conclusion, for prenatal screening of GBS from rectovaginal samples of pregnant women, our results are in accordance with CDC guidelines, which suggest using qPCR after Lim broth enrichment in addition to conventional (culture-based) detection. qPCR after Lim broth enrichment further increased the percentage of GBS-positive women, as detected by direct qPCR, from 27 to 33%, although the bacterial inoculum was low for these subjects.     

三种方法筛查孕妇B族链球菌的效果评价

目的 比较三种不同的方法检测B族链球菌的结果,对三种B族链球菌筛查方法进行效果评价.方法 收集我院2014年1月至2015年12月产科门诊孕35 ～ 37周孕妇1374例,采集孕妇的阴道拭子和肛周拭子标本,分别用血琼脂培养法,LIM肉汤增菌培养法和PCR方法检测B族链球菌.结果 1374例孕妇标本中,血琼脂培养法阳性60例,阳性检出率为4.4％,PCR方法检测阳性66例,阳性检出率为4.8％,LIM肉汤增菌培养法阳性128例,阳性检出率9.3％.三种筛查方法比较,LIM肉汤增菌培养法阳性率与PCR法,血琼脂培养法比较差异具有统计学意义.血琼脂培养法和PCR法筛查阳性率差异不具有统计学意义.以LIM肉汤增菌法为金标准,血琼脂培养法的灵敏度、特异性、阳性预测值、阴性预测值分别为46.9％,100％,100％,94.8％,PCR法的灵敏度、特异性、阳性预测值、阴性预测值分别为50％,99.6％,96.9％,95.1％.结论 LIM肉汤增菌培养法筛查B族链球菌的阳性率最高,血琼脂培养法和PCR法检测灵敏度较低,只能筛查出50％左右的GBS定植孕妇.建议临床筛查B族链球菌使用LIM肉汤增菌培养法筛查GBS,灵敏度高,特异性好.     

Use of Granada Medium To Detect Group B Streptococcal Colonization in Pregnant Women

Direct inoculation onto Granada medium (GM) in plates and tubes was compared to inoculation into a selective Todd-Hewitt broth (with 8 microg of gentamicin per ml and 15 microg of nalidixic acid per ml) for detection of group B streptococci (GBS) in pregnant women with 800 vaginal and 450 vaginoanorectal samples. Comparatively, GM was found to be as sensitive as the selective broth for the detection of GBS in vaginal specimens and more sensitive than selective broth for the detection of GBS in vaginoanorectal samples (96 versus 82%). The use of GM improved the time to reporting of a GBS-positive result by at least 24 h and reduced the direct cost of screening. We have also found that the inconvenience of anaerobic incubation of GM plates can be avoided when a cover slide is placed upon the inoculum, because aerobic incubation in GM plates with cover slides causes GBS to develop the same pigmentation that it develops with incubation under anaerobic conditions. These data support the routine use of GM plates or tubes as a more accurate, easier, and cheaper method of identification of GBS-colonized women compared to the enrichment broth technique.     

Evaluation of the New Brilliance GBS Chromogenic Medium for Screening of Streptococcus agalactiae Vaginal Colonization in Pregnant Women

Three commercial chromogenic agar media were evaluated for Streptococcus agalactiae screening in 200 vaginal swabs from pregnant women. The sensitivity and specificity were 94.3% and 100% for Granada medium (bioMérieux), 100% and 90.3% for Brilliance GBS medium (Thermo Fisher Scientific), and 100% and 98.8% for ChromID STRB medium (bioMérieux), respectively.     

Specimen storage in transport medium and detection of group B streptococci by culture

Recovery of group B streptococci (GBS) was assessed in 1,204 vaginorectal swabs stored in Amies transport medium at 4 or 21 degrees C for 1 to 4 days either by direct inoculation onto Granada agar (GA) or by culture in blood agar (BA) and GA after a selective broth enrichment (SBE) step. Following storage at 4 degrees C, GBS detection in GA was not affected after 72 h by either direct inoculation or SBE; however, GBS were not detected after SBE in the BA subculture in some samples after 48 h of storage and in GA after 96 h. After storage at 21 degrees C, loss of GBS-positive results was significant after 48 h by direct inoculation in GA and after 96 h by SBE and BA subculture; some GBS-positive samples were not detected after 24 h of storage followed by SBE and BA subculture or after 48 h of storage followed by SBE and GA subculture. Storage of swabs in transport medium, even at 4 degrees C, produced after 24 h an underestimation of the intensity of GBS colonization in most specimens. These data indicate that viability of GBS is not fully preserved by storage of vaginorectal swabs in Amies transport medium, mainly if they are not stored under refrigeration.     

Grouping and detection of group B streptococci by immunofluorescence.

Forty-nine clinical isolates of group B streptococci (GBS) were correctly grouped from broth culture by the Fluoro-Kit immunofluorescence test. A further 82 beta-haemolytic streptococci of groups A, C, D, F, and G were tested and gave no cross-reactions. The test was simple to perform and gave clear results. The Fluoro-Kit reagents, however, failed to detect GBS in 21 (51%) of 41 smears of rectal or vaginal swabs from pregnant women from which GBS were subsequently grown. Thirty-two (20%) of 159 culture-negative swabs gave positive immunofluorescent reactions.     

Modified Granada Agar Medium for the detection of group B streptococcus carriage in pregnant women

Objectives   To improve the detection rate of group B streptococci (GBS) in pregnant women, aiming at the prevention of early-onset septicemia in the newborn.
Methods   The yield from culturing two sites, vaginal and anorectal, on a Modified Granada Medium (MGM) was compared with our standard approach of culturing a vaginal swab on blood agar (BA).
Results   Samples were processed from 430 consecutive pregnant women. GBS was isolated from the vagina in 11.6% with BA, and in 13.7% with MGM. In 17.0% of anorectal samples, GBS was identified with MGM. The combination of both sites and media had a yield of 20.0%. MGM identified all but six (2%) of 310 GBS strains after aerobic incubation, with use of a cover slide, and missed only three strains (1%) after anaerobic incubation.
Conclusions   Separate culture of vaginal and anorectal samples using the same MGM agar plate resulted in an increase in detection rate for GBS of 76% as compared to BA alone. The technique is simple and results are available after overnight incubation. MGM was confirmed as a specific medium for the identification of GBS, with a sensitivity of 98–99%.     

Carriage of group B streptococcus in pregnant women from Oxford, UK

OBJECTIVE: To investigate asymptomatic vagino-rectal carriage of group B streptococcus (GBS) in pregnant women. METHODS: Women in the final trimester of pregnancy were recruited. A single vagino-rectal swab was taken, with consent, for culture of GBS. Two microbiological methods for isolation of GBS from vagino-rectal swabs were compared. The distribution of capsular serotypes of the GBS identified was determined. Epidemiological data for a subset (n = 167) of the pregnant women participating were examined. RESULTS: 21.3% were colonised vagino-rectally with GBS. Risk factors for neonatal GBS disease (maternal fever, prolonged rupture of membranes, and preterm delivery) were present in 34 of 167 women (20.4%), and the presence of these factors correlated poorly with GBS carriage. Capsular serotypes III (26.4%), IA (25.8%), V (18.9%), and IB (15.7%) were prevalent in the GBS isolates. Selective broth culture of vagino-rectal swabs was superior to selective plate culture, but the combination of both methods was associated with increased detection of GBS (7.5%). An algorithm for the identification of GBS from vagino-rectal swabs was developed. CONCLUSIONS: GBS carriage is prevalent in pregnant women in Oxfordshire, UK. The poor correlation between risk factors and GBS carriage requires further investigation in larger groups, given that the identification of these surrogate markers is recommended to guide administration of intrapartum antibiotic prophylaxis by the Royal College of Obstetricians of the UK. A selective broth culture detected more GBS carriers than a selective plate culture.     

Rapid detection of group B streptococcal colonization of the genital tract by a commercial optical immunoassay

The performance of a commercial optical immunoassay (OIA) was compared at two institutions with that of routine agar and broth culture methods for the detection of group B streptococcal (GBS) colonization of the genital tract. The Strep B OIA (Bio Star, USA) was used to test 962 vaginal swabs from pregnant women for the presence of GBS antigen. The prevalence of GBS vaginal colonization in this population was 22.4%. The OIA results were compared with those of culture on trypticase soy agar with 5% sheep blood (TSA) and broth enhanced culture (Lim broth). Sensitivity and specificity values of the OIA method compared to TSA culture alone were 82.5% and 91.8%, respectively. The sensitivity of the OIA method was equivalent to that of TSA culture (62.4% vs. 64.4%; p>0.5, ²=0.01) when the data were compared with broth culture. The extent of colonization affected the sensitivity of the OIA method: 100% of 4+, 94% of 3+, 96% of 2+, and 63% of 1+ TSA plates were detected by the OIA test. The commercial OIA method demonstrated sensitivity equivalent to that of TSA culture for the detection of GBS colonization. The OIA test offers two additional advantages over culture: reduced time required to obtain results (30 min vs. days) and the ability to detect GBS antigen in samples with compromised viability. The results of this study suggest that the Strep B OIA test can be a useful diagnostic tool in the management of early-onset GBS disease.     

Method for quantitative detection and presumptive identification of group B streptococci on primary plating

Maternal prenatal screening for group B streptococci (GBS) followed by offering of intrapartum chemoprophylaxis to carriers is one of the strategies used to reduce the incidence of neonatal early-onset GBS infections. Culturing of vaginal and anorectal swab specimens in selective broth is the screening procedure recommended by the Centers for Disease Control and Prevention. This technique is sensitive; it does not, however, allow either evaluation of the degree of colonization or detection of cocolonization with several GBS clones. We have examined the carriage rate and population dynamics of GBS in a group of Danish women during pregnancy and 1 year after delivery using a new detection method. In the present paper we describe a mixed blood agar medium (MB agar) that identifies GBS in the primary cultures by detection of a double hemolysis pattern consisting of characteristic, large zones of partial hemolysis ("CAMP zones") and of narrow zones of complete hemolysis. The MB agar was at least as sensitive as culturing in selective broth for detection of GBS in vaginal and anorectal swab specimens, and GBS strains could be identified directly on the primary plate due to the CAMP zones without the need for subculturing. The carriage rate of GBS in a group of Danish women was found to be more than 30%, a figure considerably higher than the rate that was reported previously.