Menstrual Blood as a Diagnostic Specimen for Human Papillomavirus Genotyping and Genital Tract Infection Using Next-Generation Sequencing as a Novel Diagnostic Tool
Abstract
:1. Introduction
2. Method and Materials
2.1. Patients
2.2. MB Collection and DNA Extraction
2.3. PacBio 16S rRNA HiFi Long-Read Sequencing
2.4. 16S rRNA Gene Analysis
2.5. Menstrual Blood HPV PCR
3. Results
3.1. Patients’ Characteristics
3.2. mNGS Analysis of MB
3.3. HPV DNA Detection of MB
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample ID | Number of Reads | Number of Bases (bp) | N50 Read Length (bp) | Median Read Quality (Phred) |
---|---|---|---|---|
ST04 | 551,743 | 823,059,442 | 1491 | 42.1 |
ST06 | 78,529 | 117,584,092 | 1489 | 43 |
ST08 | 370,588 | 557,280,881 | 1503 | 39.9 |
ST10 | 357,940 | 541,838,371 | 1512 | 39.1 |
ST12 | 295,312 | 446,999,092 | 1523 | 41.8 |
ST13 | 47,963 | 72,772,473 | 1532 | 43.9 |
Sample ID | Age | Cytological Diagnosis | Histological Diagnosis | HPV PCR Results in Pap Smear | HPV DNA Detected in MB | Bacterial DNA Detected in MB by mNGS |
---|---|---|---|---|---|---|
ST04 | 34 | ASC-US | CIN1 | High-risk HPV DNA detected | HPV 16 and 18 DNA detected | Sneathia (19%); Prevotella (18%); Gardnerella (7%); Fastidiosipila (6%); Megasphaera (6%); Parvimonas (5%); Atopobium (4%); Mageeibacillus (2%); Peptoniphilus (2%); Porphyromonas (2%); Dialister (2%); Gemella (2%); Peptostreptococcus (1%); Actinomycetaceae (0.7%) |
ST06 | 45 | ASC-US | CIN1/2 HPV infection | High-risk HPV DNA detected | HPV DNA not detected | Gardnerella (46%); Anaerococcus (16%); Prevotella (7%); Peptoniphilus (4%); Dialister (3%); Peptostreptococcus (2%); Atopobium (2%); Streptococcus (1%); Mycoplasmataceae (0.7%) |
ST08 | 43 | ASC-H | CIN2 | Not done | HPV DNA not detected | Gammaproteobacteria (36%); Enterobacterales (11%); Corynebacterum (6%); Fastidiosipilla (5%); Anaerococcus (0.8%); Facklamia (1%) |
ST10 | 33 | LSIL | HPV infection | Not done | High-risk HPV DNA detected (not 16 & 18) | Anaerococcus (6%); Streptococcus (5%); Lactobacillaceae (3%); Prevotellaceae (3%); Corynebacterum (2%); Peptoniphilus (1%); Prevotella (1%) |
ST12 | 39 | LSIL ASC-H | CIN3 HPV infection | High-risk HPV DNA not detected | HPV 16 DNA detected | Veillonella (33%); Peptoniphilus (6%); Streptococcus (5%); Lactobacillaceae (2%); Anaerococcus (2%); Prevotella (2%); Corynebacterum (2%); Ureaplasma (1%); Enterobacterales (1%) |
ST13 | 45 | LSIL | CIN1 HPV infection | High-risk HPV DNA detected | High-risk HPV DNA detected (not 16 and 18) | Gardnerella (37%); Lactobacillaceae (11%); Streptococcus (2%); Ureaplasma (2%); |
HPV DNA Detected in MB | HPV DNA Not Detected in MB | Sensitivity | PPV | ||
---|---|---|---|---|---|
HPV PCR using Pap smear | HPV DNA detected | 2 a | 1 b | 66.7% | 66.7% |
HPV DNA not detected | 1 c | N/A |
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Tsang, H.-F.; Cheung, Y.-S.; Yu, C.-S.A.; Chan, C.-S.S.; Wong, C.-B.T.; Yim, K.-Y.A.; Pei, X.; Wong, S.-C.C. Menstrual Blood as a Diagnostic Specimen for Human Papillomavirus Genotyping and Genital Tract Infection Using Next-Generation Sequencing as a Novel Diagnostic Tool. Diagnostics 2024, 14, 686. https://doi.org/10.3390/diagnostics14070686
Tsang H-F, Cheung Y-S, Yu C-SA, Chan C-SS, Wong C-BT, Yim K-YA, Pei X, Wong S-CC. Menstrual Blood as a Diagnostic Specimen for Human Papillomavirus Genotyping and Genital Tract Infection Using Next-Generation Sequencing as a Novel Diagnostic Tool. Diagnostics. 2024; 14(7):686. https://doi.org/10.3390/diagnostics14070686
Chicago/Turabian StyleTsang, Hin-Fung, Yui-Shing Cheung, Chi-Shing Allen Yu, Chung-Sum Sammy Chan, Chi-Bun Thomas Wong, Kay-Yuen Aldrin Yim, Xiaomeng Pei, and Sze-Chuen Cesar Wong. 2024. "Menstrual Blood as a Diagnostic Specimen for Human Papillomavirus Genotyping and Genital Tract Infection Using Next-Generation Sequencing as a Novel Diagnostic Tool" Diagnostics 14, no. 7: 686. https://doi.org/10.3390/diagnostics14070686