Tampon Specimens Research Articles

Low-coverage whole-genome sequencing for the effective diagnosis of early endometrial cancer: A pilot study

BackgroundEndometrial carcinoma (EC) is a disease that predominantly affects peri- and post-menopausal women and its incidence has continued to rise over recent years. Since the gold standard for EC diagnosis—hysteroscopic biopsy—is invasive, expensive, and unsuitable for wide use, there is an urgent need for a non-invasive method that exhibits both high sensitivity and high specificity. We therefore investigated the efficacy of UterCAD (the uterine exfoliated cell chromosomal aneuploidy detector) using tampon-collected specimens for the early detection of EC. MethodsWe prospectively recruited 51 patients with a history of abnormal bleeding and who planned to undergo hysteroscopic examination or hysterectomy between March 2020 and January 2021. Before executing an invasive procedure, a tampon was inserted into the patient's vagina for 6 h to collect exfoliated cells from the uterine cavity. Total DNA was extracted and low-coverage whole-genome sequencing was performed on an Illumina HiSeq X10, and we analyzed the differences in chromosomal status between women with EC and those bearing benign lesions using UterCAD. ResultsThirty EC patients—including 26 with endometrioid carcinoma (EEC) and four with uterine serous carcinoma (USC), as well as 14 benign cases—were enrolled in our final analysis. Copy-number variations (CNVs) were detected in tampon specimens collected from 26 EC patients (83.3%), including 21 with EEC (80.7%) and four with USC (100%). In the benign group, only one woman with focal atypical hyperplasia presented with a 10q chromosomal gain (P < 0.001). In the EC group, the most common CNVs were copy gains of 8q (N = 14), 2q (N = 4), and 10q (N = 3); and copy losses of 2q (N = 3) and 17p (N = 2). When we stratified by FIGO stage, the CNV rates in stages IA, IB, and II/III were 83.3% (15/18), 85.7% (6/7), and 80.0% (4/5), respectively. At the optimal cutoff (|Z| ≥ 2.3), UterCAD discriminated 83.3% of EC cases from benign cases, with a specificity of 92.9%. ConclusionsWe initially reported that UterCAD could serve as a non-invasive method for the early detection of EC, especially in the rare and aggressive USC subtype. The use of UterCAD might thus avoid unnecessary invasive procedures and thereby reduce the treatment burden on patients.

P-545 Efficacy, safety and acceptability of a vaginal tampon for the collection of biospecimen samples: a systematic review and meta-analysis

Abstract Study question Are self-collected vaginal tampons, as a biospecimen collection tool, comparable to current gold-standard methods in their efficacy, safety and acceptability? Summary answer Tampon efficacy for detection of gynaecological abnormalities was comparable to that of gold-standard methods in addition to being widely accepted with minimal reported adverse effects. What is known already Women’s services have poor attendance. Fear and embarrassment surrounding gynaecological health remain prevalent. Numerous long term sequalae can result from lack of gynaecological care include pelvic inflammatory disease, infertility and malignancy. Self-collection vaginal tampon samples have the potential to increase screening and diagnosis rates for multiple gynaecological conditions. Although the menstrual tampon as a biospecimen collection device has been demonstrated to be able to identify various gynaecological abnormalities, currently, possibly due to its novelty and lack of information on validity, they are not routinely used. Study design, size, duration Systematic review was conducted following PRISMA guidelines and prospectively registered with PROSPERO (CRD42022374857). A primary search through online databases was conducted, identifying 151 studies. Following this, title and abstract screening, full-text review and forward and backward citation searching were conducted in respective order, leading to a total of 92 studies for analysis. Participants/materials, setting, methods PubMed, MEDLINE, Cochrane Library, Scopus, and Web of Science were searched for eligible studies. All primary studies reporting the acceptability, efficacy or safety of vaginal tampons as a biospecimen collection device in comparison to other conventional methods of collection were included. Data was stratified based on which quantitative and/or qualitative variables were reported, baseline characteristics, and the abnormality being screened for. The Modified Newcastle Ottawa scale was used to assess risk of bias. Main results and the role of chance This systematic search identified 92 eligible studies, exploring HIV, HPV, HSV, Chlamydia trachomatis, Neisseria gonorrhoeae, cervical cancer, endometrial cancer, bacterial vaginosis, Candida albicans, Actinomyces israelii, Gardnerella vaginalis, Atopobium vaginae, and Trichomonas vaginalis. Menstrual tampons were compared to both clinician and self-collected samples including an endocervical swab, cervical brushing, cervicovaginal lavage, various tissue biopsies and urine samples. Detection of most gynaecological abnormalities did not profoundly vary between tampon collection versus gold-standard method. Self- collected tampon specimens were widely accepted with factors including lower pain scores, a greater willingness to use, convenient accessibility (especially in developing areas), and avoidance of embarrassment being reported. Very few papers reported adverse side effects and in those that did, vaginal tampon collection methods were regarded as less traumatic with a decreased likelihood of bleeding as compared to other methods including cervical swabs. Limitations, reasons for caution The effect of tampon sampling prior to other specimen collection methods could contribute to results that favoured tampon collection. A biopsy was not performed in some studies, leading to an unknown true disease status. Wider implications of the findings The use of menstrual tampons in clinical practice may help overcome barriers to accessing healthcare and participation in screening, necessary for preventing avoidable gynaecological conditions and their associated long-term complications. Further research is required to explore the optimisation of tampon specimen collection, including transportation and storage requirements. Trial registration number Not applicable

Abstract B116: Cervical HPV testing with two home self-collection methods compared to a standard clinical-collection method

Abstract Background: The purpose of this randomized trial was to examine the feasibility of using home self-sampling for HPV testing to screen for cervical cancer, as compared to HPV samples obtained by a physician or nurse practitioner during a clinical exam. Methods: Two LSU study sites were used, New Orleans and Shreveport, LA. Stratified, permuted blocks randomization was used to allocate patients to one of two methods of home sampling: a standard tampon inserted and worn for two hours, or an easy-to-use vaginal swab device (HerSwab®). All self- collected and clinical were tested for HPV 16 and 18 subtypes and other high-risk subtypes. Each self-collected sample was compared to the patient's clinical sample using McNemarâ' paired chi-squared test. Tampon and self-collected swab samples were compared using an unpaired chi-squared test. Results: A total of one hundred and seventy-four (174) eligible subjects were recruited. Ninety-five subjects were recruited from Shreveport and 79 from New Orleans. The age of the women subjects ranged from 21 to 69, with a mean age of 47 years. The predominant race was African American, 73%, with 22% white and 5% Asian, mixed race or unknown. Hispanic or Latino ethnicity was reported as 4%. Overall, 66% of home-collected specimens were returned for processing. The percent positive specimens was approximately the same, regardless of specimen source, e.g., clinical (13.5% positive), tampon (14.5% positive) or swab (15% positive). There was no statistical difference between the positive specimen rates of tampon and swab methods. By oversight, 11 clinical specimens were not collected for HPV testing. Six women had a positive self-test, but their clinical test results were not positive. All the clinical samples were sufficient for valid DNA analysis. Only one of the home swab specimens was insufficient, but 15 of the tampon specimens were insufficient. This difference is highly statistically significant, p&amp;lt; 0.0001 by Fisher's exact test. Also, as mentioned above, the return rate was better for the swab method. Fifty-five women answered the satisfaction questionnaire. Participants assigned to the tampon use complained about the procedure verbally to the study nurse, although the formal questionnaire did not reflect this. None reported problems with the swab method. Conclusion: The self-collected HPV specimens were not significantly different from the clinically collected specimens, although the tampon method had more unreturned specimens and significantly more specimens of insufficient quality for testing. Citation Format: Jerry McLarty, Donna Williams, Susan Loyd, Michael Hagensee. Cervical HPV testing with two home self-collection methods compared to a standard clinical-collection method [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr B116.

Detection of endometrial cancer via molecular analysis of DNA collected with vaginal tampons

ObjectiveWe demonstrate the feasibility of detecting EC by combining minimally-invasive specimen collection techniques with sensitive molecular testing. MethodsPrior to hysterectomy for EC or benign indications, women collected vaginal pool samples with intravaginal tampons and underwent endometrial brushing. Specimens underwent pyrosequencing for DNA methylation of genes reported to be hypermethylated in gynecologic cancers and recently identified markers discovered by profiling over 200 ECs. Methylation was evaluated individually across CpGs and averaged across genes. Differences between EC and benign endometrium (BE) were assessed using two-sample t-tests and area under the curve (AUC). ResultsThirty-eight ECs and 28 BEs were included. We evaluated 97 CpGs within 12 genes, including previously reported markers (RASSF1, HSP2A, HOXA9, CDH13, HAAO, and GTF2A1) and those identified in discovery work (ASCL2, HTR1B, NPY, HS3ST2, MME, ADCYAP1, and additional CDH13 CpG sites). Mean methylation was higher in tampon specimens from EC v. BE for 9 of 12 genes (ADCYAP1, ASCL2, CDH13, HS3ST2, HTR1B, MME, HAAO, HOXA9, and RASSF1) (all p<0.05). Among these genes, relative hypermethylation was observed in EC v. BE across CpGs. Endometrial brush and tampon results were similar. Within tampon specimens, AUC was highest for HTR1B (0.82), RASSF1 (0.75), and HOXA9 (0.74). This is the first report of HOXA9 hypermethylation in EC. ConclusionDNA hypermethylation in EC tissues can also be identified in vaginal pool DNA collected via intravaginal tampon. Identification of additional EC biomarkers and refined collection methods are needed to develop an early detection tool for EC.

Prevalence of Gardnerella vaginalis and Atopobium vaginae in Virginal Women

The objectives of this study was to determine the prevalence of Gardnerella vaginalis and Atopobium vaginae in virginal women and to establish if nonpenetrative sexual activity is associated with these organisms. The goal of this study was to assess prevalence of G. vaginalis and A. vaginae in virginal women. Forty-four virginal female patients self-collected tampon specimens, which were tested for G. vaginalis and A. vaginae by polymerase chain reaction, and provided either brief or detailed information regarding sexual practices by self-administered questionnaire. Twenty women (45%) had G. vaginalis and 3 (7%) A. vaginae detected. Among the 27 who provided detailed information, 12 (46%) had G. vaginalis and none had A. vaginae detected. G. vaginalis was significantly more likely to be detected in those who had participated in oral sex (odds ratio = 22; 95% confidence interval = 2.2-222) and hand-genital contact without penetration (P = 0.02), but not genital-genital contact. This study shows that nonpenetrative intimate sexual contact is associated with carriage of G. vaginalis in virginal women.

Comparison of a self‐administered tampon ThinPrep test with conventional pap smears for cervical cytology

To assess a self-administered tampon specimen as an alternative method of detecting cytological abnormalities and its acceptability in comparison with a conventional Papanicolou (pap) smear. Comparative observational study. Two hundred and seventeen women were recruited from the colposcopy clinic of an outer urban public teaching hospital and from sexual health clinics at suburban and major metropolitan hospital clinics. Participants inserted and immediately withdrew a tampon, then placed it into a vial of ThinPrep PreservCyt fluid. This was analysed by a local private pathology laboratory. Results were compared to a pap smear performed the same day or within the previous 6 months. All women with an abnormal result (tampon or pap smear) underwent a colposcopy, with or without biopsy as necessary. Participants completed a questionnaire after performing the tampon test. Probabilities of tampon test detecting (i) a high grade abnormality (pHG), (ii) any cervical intraepithelial neoplasia (CIN) changes (pCINany), and (iii) any abnormalities (pabn) compared to the conventional pap smear and, if abnormal, compared to the biopsy taken at colposcopy. Acceptability of the tampon test and conventional pap smear were also measured. Probabilities of the tampon test compared to pap smear: pabn sensitivity 33%, specificity 89%, PPV 59%, NPV 73%; pCINany sensitivity 23%, specificity 97%, PPV 71%, NPV 79%; pHG sensitivity 19%, specificity 98%, PPV 63%, NPV 89%. Acceptability for tampon test was 91.21% and for pap smear, 45.85%. Although the self-administered tampon ThinPrep method is a poor detector of cervical abnormalities compared to pap smear, it is highly acceptable to women. It has a relatively good negative predictive value (NPV). Our study suggests that if a more acceptable, sensitive method of cervical screening was found, which removed some of the existing barriers to conventional pap testing, screening rates for cervical cancer may improve.

Diagnosis of bacterial vaginosis on self-collected vaginal tampon specimens.

A vaginal tampon specimen was previously shown to be suitable for the molecular diagnosis of non-ulcerative sexually transmitted infections (STIs). Different tampon fluid preparations were evaluated for the diagnosis of bacterial vaginosis (BV). Women with pregnancy related problems were enrolled. Two observers evaluated the different tampon fluid preparations and vaginal smears collected during speculum examination using the Nugent score. Using the Amsel criteria, 21% of the 84 women enrolled were diagnosed with BV. Results of the tampon fluid preparations and vaginal smears showed excellent agreement for both observers (Spearman >0.80). The overall sensitivity and specificity was 91.7% (95% CI: 81.6-96.5) and 79.3% (95% CI: 67.2-87.8), respectively, using the Amsel criteria as reference standard. The tampon provides a specimen for the combined diagnosis of non-ulcerative STIs and BV. This non-invasively collected specimen may facilitate self-initiated testing and population-based studies as well as longitudinal studies that are necessary to gain insight in the epidemiology of BV related to STIs and HIV.

Estimating the prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection in indigenous women in northern Australia.

OBJECTIVE: To estimate more accurately the age specific prevalence of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, and human papillomavirus infection (HPV) in indigenous women living in urban, rural, and remote...

The tampon test for trichomoniasis: a comparison between conventional methods and a polymerase chain reaction for Trichomonas vaginalis in women.

OBJECTIVES: Trichomonas vaginalis is the most common STD worldwide and the infection has been linked with an increased risk of HIV transmission. We present a detailed comparison between conventional collection...

Patient-administered tampon-collected genital cells in the assessment of Chlamydia trachomatis infection using polymerase chain reaction.

Diagnosis of genital Chlamydia trachomatis infection in women traditionally requires a speculum examination to collect endocervical cells, followed by cell culture. This method is time consuming, requires stringent transport conditions, and is technically demanding. To compare tampons as a patient-administered collection method followed by detection with polymerase chain reaction (PCR) with the traditional endocervical swab culture followed by cell culture detection. At the emergency department of a hospital for obstetrics and gynecology, 1,000 consecutive women with symptoms suggestive of infection with C. trachomatis were tested for C. trachomatis infection by PCR on both tampon (PCR-T) and swab (PCR-S) specimen and by culture of the swab specimen. Seventeen PCR-T and 16 PCR-S specimens were positive; 16 endocervical specimens were positive by culture, and 14 of the endocervical samples were positive by the three methods. Sixty-one PCR-S samples were inadequate as shown by the lack of amplification of the beta-globin gene segment, indicating poor collection of specimens by endocervical swab for chlamydial testing. Tampon specimens collected for PCR detection provided an easy and sensitive method of detection of C. trachomatis and overcame the obstacle of endocervical sampling and subsequent stringent transport requirements of culture.

A randomized clinical trial of beta carotene vs placebo for the treatment of cervical HPV infection

The objective of the study was to determine if oral beta carotene would improve abnormalities observed in Papanicolaou smears or reduce the amount of HPV DNA in genital samples. A randomized double blind placebo controlled trial was designed for 117 women with abnormal cervical morphology, not undergoing laser ablative therapy, at The Royal Women’s Hospital, Victoria. Thirty milligrams of oral beta carotene were administered daily for 12 months. Post-intervention cervical cytology and the amount of HPV DNA present on tampon specimens as determined by polymerase chain reaction and Hybrid Capture were the main outcome measures. After 12 months therapy there was no difference between the beta carotene and placebo groups in Papanicolaou smear results (58% and 62% normal) and HPV positivity (42% and 46% positive) (P> 0.86). Women taking beta carotene were not more likely to have improved cervical cytology or a decrease in the amount of HPV DNA (P> 0.2). The median post-intervention beta carotene level was 0.63 µm ml−1 (range 0.04–1.6) for the beta carotene group and 0.15 µm ml−1 (range 0.02–1.51) for the lecithin group (P < 0.0001). This clinical trial did not identify a beneficial effect of 30 mg of oral beta carotene on cervical cytology, or on the amount of HPV DNA present from tampon specimens.

The detection of HPV DNA, the size of tampon specimens and the menstrual cycle.

To determine if HPV detection or the size of a tampon specimen is affected by the menstrual cycle. Two hundred and eighty women between 18-35 years of age attending a gynaecology clinic at The Royal Women's Hospital were enrolled. Each woman completed a questionnaire on the risk factors of HPV infection and provided a tampon specimen. Specimens were analysed for the presence of HPV DNA (polymerase chain reaction with the L1 consensus primers) after the pellet volume and number of cells was assessed. The mean age of the 298 women enrolled in this study was 27.0 years (SD 4.5, range 18-35). Ninety two (30.9%) of the tampon specimens were positive for HPV using the L1 consensus primer. The detection of HPV DNA was not associated with the quartiles of the menstrual cycle (p = 0.32). Both the pellet volume and the number of cells from a tampon specimen were greater during the mid cycle, although this was significant for the pellet volume only (p = 0.002 and 0.1 respectively). The pellet volume was not significantly associated with other variables assessed by the questionnaire. The number of cells from a tampon specimen increased with the numbers of life time sexual partners (p = 0.02) and was higher for a single marital status (p = 0.0008). The timing of the menstrual cycle effects the size of tampon specimens but not the probability of detecting HPV DNA.

The absence of genital human papillomavirus DNA in virginal women.

Our aim was to determine the prevalence of human papillomavirus (HPV) infection in virginal women and to establish if other forms of sexual activity not involving penetration may contribute to the transmission of this infection. Female patients attending medical practitioners, and female high school students were recruited. Each participant answered an anonymous self answer questionnaire and collected a tampon specimen. Tampons were analysed for the presence of human papillomavirus by the polymerase chain reaction (L1 general primer and probe). Fifty-five female participants who were of a mean age of 18 years (range 13-41) were recruited. Twenty-three reported non-penetrative sexual activity. None of the 55 women had evidence of HPV infection (0%, upper 95% confidence interval 5.4%). Twelve tampons from sexually active women with normal Papanicolaou smears and a mean age of 26 years (range 19-44) attending a hospital clinic were analysed at the same time and four were found to be positive (P = 0.0006). We conclude that HPV infection prior to sexual intercourse, as determined by tampons specimens, is rare, even in those participating in other forms of sexual activity.

Determination of menstrual prostaglandin levels in non-dysmenorrheic and dysmenorrheic subjects

We have developed a method which can measure the menstrual prostaglandin (PG) activity in a single tampon specimen by bioassays. This method makes it possible to monitor the menstrual PG activity continuously during menstruation. Using this technique, we determined the menstrual PG patterns of two normal non-dysmenorrheic subjects, one subject on oral contraceptives (OC) and one subject with moderate to severe dysmenorrhea. Two to four cycles were studied per subject. We observed three menstrual patterns among the four subjects studied. Compared to the two normal controls, the subject on OC had a significantly lower menstrual fluid total and menstrual PG activity. The mean values ± S.E. per menstrual period were 33.4 g ± 1.5 vs 21.5 g ± 2.0 and 28.6 μg (PGF 2 α equivalent) ± 1.5 vs 11.3 μg ± 4.2 respectively (control vs OC). The dysmenorrheic subject had a menstrual fluid total of 37.0 g ± 1.9 similar to the two normal controls. Her menstrual PG activity (49.8 μg ± 7.7), however, was nearly two times higher than the normal controls. In one cycle studied, the dysmenorrheic subject was treated with a PG synthetase inhibitor, ibuprofen (Motrin). Remarkable relief was achieved. The alleviation of symptoms was accompanied by a concomitant marked reduction in the menstrual PG activity.