Unveiling Hidden Risks: Intentional Molecular Screening for Sexually Transmitted Infections and Vaginosis Pathogens in Patients Who Have Been Exclusively Tested for Human Papillomavirus Genotyping
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Population and Selection Criteria
2.2. Ethical Statement
2.3. Specimen Collection
2.4. DNA Extraction
2.5. HPV Genotypes Detection
2.6. Primer Design
Pathogen | Primer Name | Gene | Amplicon Size (pb) | Gene Bank Access | Reference |
---|---|---|---|---|---|
Molluscum Contagiosum Virus | M1_MCV_F1 M1_MCV_R1 | Major envelope protein | 269 | KX214741.1 | In-house |
HSV | M1_HSV_F1 M1_HSV_R1 | DNA pol | 205 | MH697529.1 MH697449.1 | In-house |
Treponema pallidum | Primer KO Primer KO4 | DNA pol | 260 | CP073552.1 | [9] |
Haemophilus spp. | M1_Hadu_F1 M1_Hadu_R1 | 16s rRNA | 154 | LT694117.1 | In-house |
Staphylococcus aureus | M1_Stau_F1 M1_Stau_R1 | RNA pol | 123 | CP076105.1 | In-house |
Klebsiella spp. | M1_Klgr_F1 M1_Klgr_R1 | PhoE | 100 | CP073373.1 | In-house |
2.7. Positive Control Strains, Sequence Analysis, and Validation
2.8. Positive Control Constructs
2.9. PCR Assays Conditions
2.10. Statistical Analysis
3. Results
4. Discussion
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Variables | Data |
---|---|
Age (years) a | 30.2 ± 10.7 |
Gender of patients | |
Female | 220 (53.9) |
Male | 188 (46.1) |
Reason for clinical analysis b | |
HPV history | 11 (2.7) |
Condylomas | 13 (3.2) |
Early detection | 371 (90.9) |
NIC 1 | 2 (0.5) |
Genital ulcers | 2 (0.5) |
Genital warts | 9 (2.2) |
HPV Genotype | Total | Male | Female | p-Value | |||
---|---|---|---|---|---|---|---|
n | % | n | % | n | % | ||
HR-HPV | 136 | 33.3 | 62 | 15.2 | 74 | 18.1 | 0.888 |
16 | 27 | 6.6 | 13 | 3.2 | 14 | 3.4 | 0.823 |
18 | 12 | 2.9 | 7 | 1.7 | 5 | 1.2 | 0.387 |
31 | 17 | 4.2 | 8 | 2.0 | 9 | 2.2 | 0.934 |
33 | 5 | 1.2 | 2 | 0.5 | 3 | 0.7 | 0.784 |
35 | 11 | 2.7 | 6 | 1.5 | 5 | 1.2 | 0.568 |
39 | 23 | 5.6 | 10 | 2.5 | 13 | 3.2 | 0.797 |
45 | 9 | 2.2 | 5 | 1.2 | 4 | 1.0 | 0.564 |
51 | 56 | 13.7 | 23 | 5.6 | 33 | 8.1 | 0.418 |
52 | 26 | 6.4 | 10 | 2.5 | 16 | 3.9 | 0.421 |
56 | 11 | 2.7 | 2 | 0.5 | 9 | 2.2 | 0.060 |
58 | 37 | 9.1 | 12 | 2.9 | 25 | 6.1 | 0.081 |
59 | 32 | 7.8 | 13 | 3.2 | 19 | 4.7 | 0.519 |
66 | 43 | 10.5 | 14 | 3.4 | 29 | 7.1 | 0.060 |
68 | 3 | 0.7 | 1 | 0.2 | 2 | 0.5 | 0.657 |
LR HPV | 153 | 37.5 | 90 | 22.1 | 63 | 15.4 | 0.000 |
6 | 122 | 29.9 | 74 | 18.1 | 48 | 11.8 | 0.000 * |
11 | 63 | 15.4 | 41 | 10.0 | 22 | 5.4 | 0.001 * |
Pathogens | Total | Male | Female | p-Value | |||
---|---|---|---|---|---|---|---|
n | % | n | % | n | % | ||
Candida spp. | 48 | 11.8 | 27 | 6.6 | 21 | 5.1 | 0.132 |
Ureaplasma spp. | 101 | 24.8 | 36 | 8.8 | 65 | 15.9 | 0.015 * |
Trichomonas vaginalis | 1 | 0.02 | 0 | 0.0 | 1 | 0.2 | 0.355 |
Neisseria gonorrhoeae | 1 | 0.2 | 2 | 0.5 | 3 | 0.7 | 0.657 |
Chlamydia trachomatis | 11 | 2.7 | 5 | 1.2 | 6 | 1.5 | 0.966 |
Mycoplasma spp. | 28 | 6.9 | 5 | 1.2 | 23 | 5.6 | 0.002 * |
Treponema pallidum | 0 | 0.0 | 0 | 0.0 | 0 | 0.0 | - |
Haemophilus spp. | 132 | 32.4 | 69 | 16.9 | 63 | 15.4 | 0.083 |
Staphylococcus aureus | 29 | 7.1 | 12 | 2.9 | 17 | 4.2 | 0.598 |
Klebsiella spp. | 16 | 3.9 | 5 | 1.2 | 11 | 2.7 | 0.225 |
HSV 1/2 | 4 | 1.0 | 0 | 0.0 | 4 | 1.0 | 0.063 |
MCV | 13 | 3.2 | 10 | 2.5 | 3 | 0.7 | 0.023 * |
HPV Results | Total | STI Positive | STI Negative | OR | 95% CI | p-Value | |||
---|---|---|---|---|---|---|---|---|---|
n | % | n | % | n | % | ||||
HPV Positive Test | |||||||||
Yes | 294 | 72.1 | 179 | 43.9 | 115 | 28.2 | 1.053 | 0.677–1.637 | 0.819 |
No | 114 | 27.9 | 68 | 16.7 | 46 | 11.3 | - | - | |
HPV multiple infection | |||||||||
Yes | 110 | 27.0 | 65 | 15.9 | 45 | 11.0 | 0.921 | 0.590–1.438 | 0.716 |
No | 182 | 44.6 | 116 | 28.4 | 298 | 73.0 | - | - |
Anatomical Site | Total | Male | Female | HPV+/STI+ | ||||
---|---|---|---|---|---|---|---|---|
n | % | n | % | n | % | n | % | |
External genitalia * | 19 | 4.7 | 9 | 4.8 | 10 | 4.5 | 14 | 3.4 |
wart | 26 | 6.4 | 24 | 12.8 | 2 | 0.9 | 14 | 3.4 |
Urethra | 86 | 21.1 | 84 | 44.7 | 2 | 0.9 | 49 | 12.0 |
BG | 7 | 1.7 | 7 | 1.7 | - | - | 2 | 0.5 |
Foreskin | 3 | 0.7 | 3 | 0.7 | - | - | 3 | 0.7 |
Penis | 54 | 13.2 | 54 | 13.2 | - | - | 36 | 8.8 |
Anus | 4 | 1.0 | 4 | 1.0 | 0 | 0.0 | 2 | 0.5 |
Vagina | 3 | 0.7 | - | - | 3 | 0.7 | 3 | 0.7 |
Cervix | 206 | 50.5 | - | - | 206 | 50.5 | 124 | 30.4 |
Pathogens | Total | HPV Positive | HPV Negative | OR | 95% CI | p-Value * | |||
---|---|---|---|---|---|---|---|---|---|
n | % | n | % | n | % | ||||
Candida spp. | 48 | 11.8 | 40 | 9.8 | 8 | 2.0 | 2.087 | 0.945–4.608 | 0.064 |
Ureaplasma spp. | 101 | 24.8 | 66 | 16.2 | 35 | 8.6 | 0.653 | 0.403–1.059 | 0.083 |
T. vaginalis | 1 | 0.2 | 1 | 0.2 | 0 | 0.0 | 1.389 | 1.307–1.476 | 0.553 |
N. gonorrhoeae | 3 | 0.7 | 2 | 0.5 | 1 | 0.2 | 0.774 | 0.069–8.620 | 0.835 |
C. trachomatis | 11 | 2.7 | 6 | 1.5 | 5 | 1.2 | 0.454 | 0.136–1.519 | 0.189 |
Mycoplasma spp. | 28 | 6.9 | 25 | 6.1 | 3 | 0.7 | 3.439 | 1.170–11.62 | 0.035 |
Haemophilus spp. | 132 | 32.4 | 91 | 22.3 | 41 | 10.0 | 0.798 | 0.506–1.259 | 0.331 |
S. aureus | 29 | 7.1 | 27 | 6.6 | 2 | 0.5 | 5.663 | 1.324–24.21 | 0.009 |
Klebsiella spp. | 16 | 3.9 | 12 | 2.9 | 4 | 1.0 | 1.170 | 0.369–3.706 | 0.789 |
HSV 1/2 | 4 | 1.0 | 4 | 1.0 | 0 | 0.0 | 1.393 | 1.310–1.481 | 0.211 |
MCV | 13 | 3.2 | 13 | 3.2 | 0 | 0.0 | 1.406 | 1.320–1.497 | 0.022 |
HPV Genotype | Total | STI Positive | STI Negative | OR | CI 95% | p-Value | |||
---|---|---|---|---|---|---|---|---|---|
n | % | n | % | n | % | ||||
HR-HPV | 136 | 33.3 | 80 | 19.6 | 56 | 13.7 | 0.898 | 0.590–1.367 | 0.616 |
16 | 27 | 6.6 | 13 | 3.2 | 14 | 3.4 | 0.583 | 0.267–1.276 | 0.173 |
39 | 23 | 5.6 | 15 | 3.7 | 8 | 2.0 | 1.237 | 0.512–2.987 | 0.800 |
51 | 56 | 13.7 | 36 | 8.8 | 20 | 4.9 | 1.203 | 0.669–2.163 | 0.537 |
52 | 26 | 6.4 | 14 | 3.4 | 12 | 2.9 | 0.746 | 0.336–1.657 | 0.471 |
58 | 37 | 9.1 | 23 | 5.6 | 14 | 3.4 | 1.078 | 0.537–2.163 | 0.832 |
59 | 32 | 7.8 | 21 | 5.1 | 11 | 2.7 | 1.267 | 0.594–2.704 | 0.540 |
66 | 43 | 10.5 | 29 | 7.1 | 14 | 3.4 | 1.397 | 0.714–2.733 | 0.328 |
LR HPV | 153 | 37.5 | 98 | 24.0 | 55 | 13.5 | 1.268 | 0.838–1.917 | 0.261 |
6 | 122 | 29.9 | 74 | 18.1 | 48 | 11.8 | 1.007 | 0.653–1.554 | 0.975 |
11 | 63 | 15.4 | 41 | 10.0 | 22 | 5.4 | 1.258 | 0.718–2.203 | 0.423 |
HPV Results | STI Results | STI Results | OR | CI 95% | p-Value |
---|---|---|---|---|---|
HPV 16 | |||||
Genotype 16 | HSV Positive | HSV Negative | OR | CI 95% | p-Value |
Positive | 2 (0.5) | 25 (6.1) | 15.160 | 2.049–112.165 | 0.000 |
Negative | 2 (0.5) | 379 (92.9) | - | - | |
HPV 16 | |||||
Genotype 16 | Ureaplasma spp. Positive | Ureaplasma spp. Negative | OR | CI 95% | p-Value |
Positive | 2 (0.5) | 25 (6.1) | 0.228 | 0.053–0.980 | 0.031 |
Negative | 99 (24.3) | 282 (69.1) | - | - | |
HPV 6 | |||||
Genotype 6 | MCV Positive | MCV Negative | OR | CI 95% | p-Value |
Positive | 10 (2.5) | 112 (27.5) | 8.423 | 2.276–31.173 | 0.000 |
Negative | 3 (0.7) | 283 (69.4) | - | - | |
HPV 6 | |||||
Genotype 6 | HSV Positive | HSV Negative | OR | CI 95% | p-Value |
Positive | 3 (0.7) | 119 (29.2) | 7.185 | 0.740–69.772 | 0.048 |
Negative | 1 (0.2) | 285 (69.9) | - | - | |
HPV 66 | |||||
Genotype 66 | HSV Positive | HSV Negative | OR | CI 95% | p-Value |
Positive | 2 (0.5) | 41 (10.0) | 8.854 | 1.215–64.536 | 0.010 |
Negative | 2 (0.5) | 363 (89.0) | - | - |
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Hernández-Rosas, F.; Rey-Barrera, M.; Hernández-Barajas, F.; Rangel-Soto, C.; García-González, M.S.; Franco-González, S.S.; de León-Bautista, M.P. Unveiling Hidden Risks: Intentional Molecular Screening for Sexually Transmitted Infections and Vaginosis Pathogens in Patients Who Have Been Exclusively Tested for Human Papillomavirus Genotyping. Microorganisms 2023, 11, 2661. https://doi.org/10.3390/microorganisms11112661
Hernández-Rosas F, Rey-Barrera M, Hernández-Barajas F, Rangel-Soto C, García-González MS, Franco-González SS, de León-Bautista MP. Unveiling Hidden Risks: Intentional Molecular Screening for Sexually Transmitted Infections and Vaginosis Pathogens in Patients Who Have Been Exclusively Tested for Human Papillomavirus Genotyping. Microorganisms. 2023; 11(11):2661. https://doi.org/10.3390/microorganisms11112661
Chicago/Turabian StyleHernández-Rosas, Fabiola, Manuel Rey-Barrera, Flavio Hernández-Barajas, Claudia Rangel-Soto, Mariana Socorro García-González, Shumeyker Susmith Franco-González, and Mercedes Piedad de León-Bautista. 2023. "Unveiling Hidden Risks: Intentional Molecular Screening for Sexually Transmitted Infections and Vaginosis Pathogens in Patients Who Have Been Exclusively Tested for Human Papillomavirus Genotyping" Microorganisms 11, no. 11: 2661. https://doi.org/10.3390/microorganisms11112661