Extended Data Fig. 2: Nested PCR-qPCR assay to detect the 16S GBS gene. | Nature Microbiology

Extended Data Fig. 2: Nested PCR-qPCR assay to detect the 16S GBS gene.

From: Placental Streptococcus agalactiae DNA is associated with neonatal unit admission and foetal pro-inflammatory cytokines in term infants

Extended Data Fig. 2: Nested PCR-qPCR assay to detect the 16S GBS gene.

a, Schematic diagram of the nested PCR-qPCR assay. b, Multiple sequence alignment of the amplicons generated with the nested PCR-qPCR assay and the 16S GBS gene sequence (SnapGene Viewer 6.1). The sequences of the PCR amplicons (n = 6 from 4 placentas positive for 16S and sip, and 2 placentas positive for 16S) were determined by Sanger sequencing using the second-round (inner) reverse and forward primers. This analysis confirmed that the sequence of the qPCR products perfectly aligned to the GBS 16S gene sequence. c, Multiple sequence alignment of the 16S gene from various Streptococci: S. agalactiae (GBS) (NR_040821.1), S. dysgalactiae (MH393517.1), S. salivarius (KM221948.1), S. anginosus (MF578782.1), S. vestibularis (NR_042777.1), S. pyogenes (NR_028598.1), S. suis (NR_115737.1), S. urinalis (NR_115738.1), S. equi (NR_116010.1), S. uberis (U41048.1). The position of the inner primers, outer primers and Taqman probe of the nested PCR-qPCR assay are indicated. The red frames highlight regions that differ between species amplified using the qPCR inner primers.

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