Molecular and Serological Diagnostic Approach to Define the Microbiological Origin of Blood Culture-Negative Infective Endocarditis
Abstract
:1. Introduction
2. Patients and Methods
2.1. Study Design
Data Collection
2.2. Blood Sample Collection
2.3. Processing of Valve Tissue Sections from BCNIE Patients
2.4. Direct Smears and Culture of Valve Sections
2.5. Molecular Techniques
2.6. DNA Concentration and Purity
2.7. DNA Cycle Sequencing
2.8. Bioinformatics for Sequence Interpretation and Statistical Analysis
2.9. Serological Testing
3. Results
4. Discussion
Study Limitations
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
BCNIE | Blood culture-negative infective endocarditis |
IE | Infective endocarditis |
PCR | Polymerase chain reaction |
KOH | Potassium hydroxide |
SDA | Sabouraud’s dextrose agar |
GM | Aspergillus galactomannan antigen |
References
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Patient No. | Organisms Identified by Valve Broad-Range PCR and Direct Sequencing (Number) | Valve Culture | Blood Broad-Range PCR and Direct Sequencing | Histopathology |
---|---|---|---|---|
1 | Staphylococcus aureus | Positive | Positive | typical IE |
2 | Staphylococcus epidermidis | Positive | Positive | typical IE |
3 | Streptococcus mitis | Negative | Negative | typical IE |
4 | Brucella spp. | Negative | Negative | typical IE |
5 | Streptococcus mutans | Negative | positive | typical IE |
6 | Aspergillus flavus | Negative | Negative | typical IE |
7 | Streptococcus mitis | It was positive for Streptococci spp. | Negative | typical IE |
8 | Enterococcus faecalis | It was positive for Enterococci spp. | Negative | typical IE |
9 | Streptococcus oralis | Negative | Negative | typical IE |
10 | Streptococcus mitis | Negative | Negative | typical IE |
11 | Bartonella henselae | Negative | Negative | typical IE |
12 | Enterococcus faecalis | Negative | Negative | typical IE |
13 | Streptococcus agalactiae | Negative | Positive | typical IE |
14 | Streptococcus anginosus | It was positive for Viridans Streptococci | Negative | typical IE |
15 | Acinetobacter baumannii | It was positive for Pseudomonas aeruginosa | Negative | typical IE |
16 | Streptococcus mutans | Negative | Negative | typical IE |
17 | Haemophilus parainfluenzae | Negative | Negative | typical IE |
18 | Candida albicans (1) | Positive | Positive | typical IE |
19 | Staphylococcus aureus | Negative | Negative | typical IE |
20 | Staphylococcus epidermidis | Negative | Negative | typical IE |
21 | Streptococcus oralis | Negative | Negative | typical IE |
22 | Aspergillus flavus | Negative | Negative | typical IE with dichotomously branched septate fungal hyphae on H&E |
23 | Staphylococcus aureus | Positive | Positive | typical IE |
24 | Streptococcus mutans | Negative | Negative | typical IE |
25 | Staphylococcus aureus | Negative | Negative | typical IE |
26 | Brucella spp. | Negative | Negative | typical IE |
27 | Staphylococcus epidermidis | Negative | Positive | typical IE |
28 | Staphylococcus aureus | Positive | Positive | typical IE |
29 | Brucella spp. | Negative | Negative | typical IE |
30 | Aspergillus flavus | Positive | Positive | typical IE with dichotomously branched septate fungal hyphae on H&E |
Pathogen (Number) | Blood Broad-Range PCR and Direct Sequencing | Valve Culture | Valve Broad-Range PCR and Direct Sequencing | Serology |
---|---|---|---|---|
Streptococcus spp.(10) | 2 | 2 | 10 | 1 positive for GM |
Staphylococcus aureus (6) | 4 | 3 | 5 | |
Aspergillus flavus (4) | 2 | 1 | 3 | 3 positive for GM |
Staphylococcus epidermidis (3) | 2 | 1 | 3 | 1 positive for GM |
Brucella spp. (4) | - | - | 3 | 4 positive for anti-Brucella antibodies |
Enterococcus faecalis (2) | - | 1 | 2 | |
Bartonella henselae (2) | - | - | 1 | 2 positive for anti-Bartonella henselae antibodies |
Acinetobacter baumannii (1) | - | Contamination (Pseudomonas aeruginosa) | 1 | |
Haemophilus parainfluenzae (1) | - | - | 1 | |
Candida albicans (1) | 1 | 1 | 1 | 1 positive for GM |
Coxiella burnetii (1) | - | - | - | 1 positive for anti-Coxiella burnetii antibodies |
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El-Ashry, A.H.; Saad, K.; Obiedallah, A.A.; Elhoufey, A.; Dailah, H.G.; Hussein, M.S.A. Molecular and Serological Diagnostic Approach to Define the Microbiological Origin of Blood Culture-Negative Infective Endocarditis. Pathogens 2022, 11, 1220. https://doi.org/10.3390/pathogens11111220
El-Ashry AH, Saad K, Obiedallah AA, Elhoufey A, Dailah HG, Hussein MSA. Molecular and Serological Diagnostic Approach to Define the Microbiological Origin of Blood Culture-Negative Infective Endocarditis. Pathogens. 2022; 11(11):1220. https://doi.org/10.3390/pathogens11111220
Chicago/Turabian StyleEl-Ashry, Amira H., Khaled Saad, Ahmed A. Obiedallah, Amira Elhoufey, Hamad Ghaleb Dailah, and Mohammed Salah A. Hussein. 2022. "Molecular and Serological Diagnostic Approach to Define the Microbiological Origin of Blood Culture-Negative Infective Endocarditis" Pathogens 11, no. 11: 1220. https://doi.org/10.3390/pathogens11111220