The Pathogenesis Of Streptococcus anginosus In Aerobic Vaginitis

Figures & data

Table 1 Primers Used In This Study

Name	Sequence (5ʹ–3ʹ)	Target	Use
27Fʹ	AGRGTTYGATYCTGGCTCAG	16S rRNA V1-V2	Sequencing
338Rʹ	GCTGCCTCCCGTAGGAGT	16S rRNA V1-V2	Sequencing
Up-F1	AGAAACTTTCCTGAGCCAATTGCTAGG	Upstream fragment of sag	Construction of mutant
Up-R1	ATCTCCCATTATATCTAGTTTGTCCTCCTTATAAAATGATGTGTAATATTACAA	Upstream fragment of sag	Construction of mutant
erm-F2	ATAAGGAGGACAAACTAGATATAATGGGAGATAAGACGGTTCGTGTT	erm	Construction of mutant
erm-R2	AACTAGAAAATACAATGCGACTCATAGAATTATTTCCTCCCGT	erm	Construction of mutant
Down-F3	TAATTCTATGAGTCGCATTGTATTTTCTAGTTGGATGTTAAAACCCATTC	Downstream fragment of sag	Construction of mutant
Down-R3	AGTGCTGATAATATTTGTGTTCGAATACTTTCC	Downstream fragment of sag	Construction of mutant
sag-F-primer	ATTACACATCATTTTATAAGGAGGACAAACTATGTTAA	sag fragment	Verification of sag cluster
sag-R-primer	TTCTCCTCATCTATCGTTTCATTTTCAGAC	sag fragment	Verification of sag cluster

Figure 1 The PCR fragments for the knockout of the sag gene and the S. anginosus (ATCC33397/NCTC10713) growing on Blood Agar Plate (BAP). (A) The up+erm+down fragment after fusion PCR; (B) is the up+erm fragment and (C) is the erm+down fragment. (D) The sag fragment of ATCC33397 (WT); (E and F) are erm fragments; (G) is upstream and (H) is downstream of the sag coding area. (I) The sag fragment of the mutant amplified from Δsag ATCC33397 with primers sag-F-primer and sag-R-primer shown in Table 1. (J) The wild type, withαhaemolytic activity, and (K) is the mutant with no sag gene and no haemolytic activity.

Table 2 The Symptoms Of Each Patient With AV. “1” Represents Yes, Meaning The Patient Had The Characteristic, And “0” Represents No

ID	Predominance	Purulent Discharge	Itching	Vaginal Wall Reddening	Dyspareunia	Rotten Odour
AV1	S. anginosus	1	1	0	0	1
AV2	S. anginosus	1	1	1	0	0
AV3	S. anginosus	1	1	1	0	0
AV4	L. inners	0	1	0	0	0
AV5	Atopobium. vaginae	1	0	0	0	1
AV6	S. anginosus	1	0	0	0	0
AV7	S. anginosus	1	1	1	1	0
AV8	Aerococcus. christensenii	1	1	1	1	0
AV9	S. anginosus	1	1	0	0	0
AV10	S. anginosus	1	0	0	0	0

Figure 2 The clustering analysis of AV, BV and NM. The relevance of species to each sample is shown. The deeper the colour is, the more relevant the sample is to the species.

Figure 3 The dendrograms of AV, BV and NM samples. The dimensions of the tree show the distance of the samples.

Figure 4 The average abundance of genera in AV, NM and BV samples. The total abundance of each group is 1.

Figure 5 Comparison of the abundance of Streptococcus between AV, NM and BV samples. **p<0.01.

Figure 6 The average abundance of species in AV, NM and BV samples. The total abundance of each group is 1.

Figure 7 The average abundance of each species in the AV microbiome. S. anginosus was significantly more abundant than other species. **p<0.01.

Figure 8 The growth curves of S. anginosus (ATCC33397) and the Δsag mutant.

Figure 9 The cytotoxicity of S. anginosus (ATCC33397) and the Δsag mutant was significantly different. **p<0.01.