Direct identification of Streptococcus agalactiae and capsular type by real-time PCR in vaginal swabs from pregnant women

Objectives: Application of SYBR Green real-time PCR and bacterial isolation methods to detect Streptococcus agalactiae (GBS) carriage in women at 35-37 weeks’ gestation. Patients and Method: Use of SYBR Green real-time PCR and bacterial isolation methods to detect GBS in 116 women at 35 - 37 weeks’ gestation. Results: The rate of carrier of GBS in women at 35 - 37 weeks gestation was 9.5% (11 pregnant women), in which real-time PCR method was positive in all positive GBS women, while bacterial culture method was positive at 6% (7 pregnant women). These methods (culture and real-time PCR) had substantial agreement with the value of Kappa is 0,77. Checking for the targeted sequence amplification of real-time PCR by the curve of melting temperature and agarose electrophoresis of amplified product, the real-time PCR product was correctly targeted at the expected genetic sequence. Conclusion: SYBR Green real-time PCR method for detecting GBS in pregnant women is useful, low-cost and easy for performing. Therefore, it is suitable for detecting GBS in diagnostic laboratories where real-time PCRs are available. Key words: Streptococcus agalactiae (GBS), real-time PCR, pregnant woman

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Development and analytical validation of real-time PCR for the detection of Streptococcus agalactiae in pregnant women

BMC Pregnancy and Childbirth ◽

10.1186/s12884-020-03038-z ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Daniel F. Escobar ◽

Diego A. Diaz-Dinamarca ◽

Carlos F. Hernández ◽

Daniel A. Soto ◽

Ricardo A. Manzo ◽

...

Keyword(s):

Pregnant Women ◽

Real Time ◽

Streptococcus Agalactiae ◽

Real Time Pcr ◽

Analytical Validation

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Diagnosis of Toxoplasmosis in Pregnant Women Using Enzyme Immunoassay and Nested Real Time PCR in Al-Najaf city/Iraq

International Journal of Pharmaceutical Research ◽

10.31838/ijpr/2020.12.02.0022 ◽

2020 ◽

Vol 12 (02) ◽

Keyword(s):

Pregnant Women ◽

Enzyme Immunoassay ◽

Real Time ◽

Real Time Pcr

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Two Reliable Methodical Approaches for Non-Invasive RHD Genotyping of a Fetus from Maternal Plasma

Diagnostics ◽

10.3390/diagnostics10080564 ◽

2020 ◽

Vol 10 (8) ◽

pp. 564

Author(s):

Jana Bohmova ◽

Marek Lubusky ◽

Iva Holuskova ◽

Martina Studnickova ◽

Romana Kratochvilova ◽

...

Keyword(s):

Pregnant Women ◽

Real Time ◽

Real Time Pcr ◽

Dna Analysis ◽

Methodological Approach ◽

Maternal Plasma ◽

Buccal Swab ◽

Non Invasive ◽

Pcr Method ◽

Parallel Reactions

Noninvasive fetal RHD genotyping is an important tool for predicting RhD incompatibility between a pregnant woman and a fetus. This study aimed to assess a methodological approach other than the commonly used one for noninvasive fetal RHD genotyping on a representative set of RhD-negative pregnant women. The methodology must be accurate, reliable, and broadly available for implementation into routine clinical practice. A total of 337 RhD-negative pregnant women from the Czech Republic region were tested in this study. The fetal RHD genotype was assessed using two methods: real-time PCR and endpoint quantitative fluorescent (QF) PCR. We used exon-7-specific primers from the RHD gene, along with internal controls. Plasma samples were analyzed and measured in four/two parallel reactions to determine the accuracy of the RHD genotyping. The RHD genotype was verified using DNA analysis from a newborn buccal swab. Both methods showed an excellent ability to predict the RHD genotype. Real-time PCR achieved its greatest accuracy of 98.6% (97.1% sensitivity and 100% specificity (95% CI)) if all four PCRs were positive/negative. The QF PCR method also achieved its greatest accuracy of 99.4% (100% sensitivity and 98.6% specificity (95% CI)) if all the measurements were positive/negative. Both real-time PCR and QF PCR were reliable methods for precisely assessing the fetal RHD allele from the plasma of RhD-negative pregnant women.

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