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Research Article
Molecular characteristics of Brucella melitensis isolates from humans in Qinghai Province, China
Liqing Xu
qinghaiinstitute for endemic disease prevention and control
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
The prevalence of human brucellosis in Qinghai Province has been increasing rapidly, with confirmed cases distributed across 31 counties. However, the epidemiology of brucellosis transmission has not been fully elucidated. To characterize the two isolated strains, multiple locus variable-number tandem repeats analysis (MLVA) and whole-genome single-nucleotide polymorphism (SNP)-based approaches were employed.
Methods
Blood samples were isolated from two males that were confirmed B. melitensis positive following MLVA. Genomic DNA was extracted from these samples, and whole-genome sequencing (WGS) was performed. Next, SNP-based phylogenetic analysis was performed to compare the two strains to 94 B. melitensis strains (complete genome and draft genome) retrieved from online databases.
Results
The two Brucella isolates were identified as B. melitensis biovar 3 (QH2019001 and QH2019005) following conventional biotyping and were found to have differences in their variable number tandem repeats (VNTRs) using MLVA-16. Phylogenetic examination assigned the 96 strains to five genotype groups, with QH2019001 and QH2019005 assigned to the same group, but different subgroups. Moreover, the QH2019005 strain was assigned to a new subgenotype, IIj, within genotype II. These findings were then combined to determine the geographic origin of the two Brucella strains.
Conclusions
Utilizing a whole-genome SNP-based approach enabled differences between the two B. melitensis strains to be more clearly resolved, and facilitated the elucidation of their different evolutionary histories. This approach also revealed that QH2019005 is a member of a new subgenotype (IIj) with an ancient origin in the eastern Mediterranean Sea.
Keywords
Brucella melitensis, MLVA, whole-genome sequencing, SNP
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Peer Review Timeline
CURRENT STATUS: Under Revision
Version (no preprint)
Posted 16 Dec, 2020
Editorial decision: Major revision
On 16 Dec, 2020
Review #2 received
Received 13 Dec, 2020
Review #1 received
Received 07 Dec, 2020
Reviewer #2 agreed
On 03 Dec, 2020
Reviewers invited
Invitations sent on 02 Dec, 2020
Reviewer #1 agreed
On 02 Dec, 2020
Editor assigned
On 30 Nov, 2020
Submission checks complete
On 30 Nov, 2020
Editor invited
On 30 Nov, 2020
This version was not preprinted
Version 1
Posted 28 Sep, 2020
No community comments so far
Review #2 received
Received 10 Nov, 2020
Editorial decision: Major revision
On 10 Nov, 2020
Reviewer #3 agreed
On 09 Nov, 2020
Reviewer #2 agreed
On 22 Oct, 2020
Reviewer #1 agreed
On 20 Oct, 2020
Review #1 received
Received 20 Oct, 2020
Reviewers invited
Invitations sent on 29 Sep, 2020
Editor assigned
On 28 Sep, 2020
Editor invited
On 27 Sep, 2020
Submission checks complete
On 24 Sep, 2020
First submitted
On 22 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
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This preprint is under consideration at Infectious Diseases of Poverty. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Molecular characteristics of Brucella melitensis isolates from humans in Qinghai Province, China
Liqing Xu
qinghaiinstitute for endemic disease prevention and control
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Revision
Version (no preprint)
Posted 16 Dec, 2020
Editorial decision: Major revision
On 16 Dec, 2020
Review #2 received
Received 13 Dec, 2020
Review #1 received
Received 07 Dec, 2020
Reviewer #2 agreed
On 03 Dec, 2020
Reviewers invited
Invitations sent on 02 Dec, 2020
Reviewer #1 agreed
On 02 Dec, 2020
Editor assigned
On 30 Nov, 2020
Submission checks complete
On 30 Nov, 2020
Editor invited
On 30 Nov, 2020
This version was not preprinted
Version 1
Posted 28 Sep, 2020
No community comments so far
Review #2 received
Received 10 Nov, 2020
Editorial decision: Major revision
On 10 Nov, 2020
Reviewer #3 agreed
On 09 Nov, 2020
Reviewer #2 agreed
On 22 Oct, 2020
Reviewer #1 agreed
On 20 Oct, 2020
Review #1 received
Received 20 Oct, 2020
Reviewers invited
Invitations sent on 29 Sep, 2020
Editor assigned
On 28 Sep, 2020
Editor invited
On 27 Sep, 2020
Submission checks complete
On 24 Sep, 2020
First submitted
On 22 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
The prevalence of human brucellosis in Qinghai Province has been increasing rapidly, with confirmed cases distributed across 31 counties. However, the epidemiology of brucellosis transmission has not been fully elucidated. To characterize the two isolated strains, multiple locus variable-number tandem repeats analysis (MLVA) and whole-genome single-nucleotide polymorphism (SNP)-based approaches were employed.
Methods
Blood samples were isolated from two males that were confirmed B. melitensis positive following MLVA. Genomic DNA was extracted from these samples, and whole-genome sequencing (WGS) was performed. Next, SNP-based phylogenetic analysis was performed to compare the two strains to 94 B. melitensis strains (complete genome and draft genome) retrieved from online databases.
Results
The two Brucella isolates were identified as B. melitensis biovar 3 (QH2019001 and QH2019005) following conventional biotyping and were found to have differences in their variable number tandem repeats (VNTRs) using MLVA-16. Phylogenetic examination assigned the 96 strains to five genotype groups, with QH2019001 and QH2019005 assigned to the same group, but different subgroups. Moreover, the QH2019005 strain was assigned to a new subgenotype, IIj, within genotype II. These findings were then combined to determine the geographic origin of the two Brucella strains.
Conclusions
Utilizing a whole-genome SNP-based approach enabled differences between the two B. melitensis strains to be more clearly resolved, and facilitated the elucidation of their different evolutionary histories. This approach also revealed that QH2019005 is a member of a new subgenotype (IIj) with an ancient origin in the eastern Mediterranean Sea.
Figure 1
Figure 2
Figure 3