The Development of a Rapid Vaginitis Test Kit
Maya Rao, Secondary Division, 69118, William Mason High School, Ohio
Research Question Results
Is there a need for an at-home rapid vaginitis test and if so, is it possible to create it?
Acknowledgements
The author would like to thank Carol Anderson for her inspiration and continuous support of this project.
A special thanks to Victoria Rao and Alexander Chernyakhovsky for assistance with editing writings
and electrical circuit model creation, respectively.
Abstract
Motivation: The Mayo Clinic defines vaginitis as the inflammation of the vagina that can result in
discharge, itching, and pain. Bacterial Vaginosis and Vaginal Candidiasis, or more commonly known as
vaginal yeast infection, are responsible for 90% of cases of vaginitis [7]. An estimated 75% of women
will get a yeast infection during their lifetime [5], and 33% of women will get bacterial vaginosis.
Diagnosing the infection can be challenging due to late-onset symptoms or asymptomatic infection. If
left untreated, a yeast infection can lead to serious health problems [3]. There is currently no accurate
vaginitis test kit on the market, thereby necessitating lab-based testing resulting in treatment delays. In
case of bacterial infection, antibiotics often need to be prescribed by a doctor.
Objective: There are two primary objectives of this study. The first is to determine the need for an
at-home rapid vaginitis test. The second is to research the methods that can be used to create an
easily accessible and effective test.
Methods: This study uses survey-based scientific methods to determine the motivation for the creation
of an at-home rapid vaginitis test kit. To create the rapid test kit, background technical research was
conducted to design a theoretical working product prototype.
Findings: Survey results support the need for an at-home test kit (59.7%) and further education
regarding yeast infections (87.5%). A rapid test can be created by testing for the presence of the
enzyme sialidase. Research published in Nature found a biochemiluminescent sialidase assay that
uses a substrate derivatized with firefly luciferin that can accurately detect the presence of sialidase
[8]. The one-step assay enables a compact and easy-to-use product to be designed to create a rapid
vaginitis test.
Summary: This study supplies the necessary ingredients, models, and instructions required to
manufacture an accessible and effective rapid vaginitis test kit.
Methods
Survey Methods
• Target population: The target population of the survey was biomedical science high school students
ages 13-19.
• Data collection: The data was collected via a self-administered survey that included demographic
questions regarding age and biological sex as well as questions to gain an understanding of the
students’ perceptions of yeast infections.
• The survey was approved by the Biomedical Sciences program at William Mason High School. The
fully anonymized web-based survey was administered to biomedical science students for 7 days.
Participation was voluntary and no incentives were provided to the participants.
• Data Analysis: The survey data was automatically analyzed by Google Forms which provided the
summary statistics.
• Data sharing: Data was fully anonymized and only summary statistics and charts were used to
share results. Statistics, pie charts, and bar graphs from the survey were utilized in informational
pamphlets and posters.
• Biases: Only biomedical sciences students participated in this survey. This may have skewed the
results of respondents’ overall knowledge of yeast infections. The survey had a majority of female
participants (77.8%).
Prototype Development Methods
• Target population: The target population of the rapid at-home test is female teenagers and adults.
The test could also be used by medical providers as a rapid diagnostic tool.
• Prototype Design: The product was designed using the well-established engineering design
process [4]. The primary steps of the design process include defining a problem, generating
concepts, developing a solution, constructing, testing, and evaluating the solution.
• Background research to justify the necessity of a rapid test was conducted followed by research to
create a working product. First, a protocol for an assay was extracted [8]. Next, a prototype mockup
was created as a 2D sketch. Based on the initial sketch, 3D model prototypes were designed,
3D-printed, and tested, to ensure functionality. The digital rendition of the final prototype design is
seen in Figure 2.
• Possible Errors: The prototype implementation could not be fully tested due to the inaccessibility of
the assay ingredients to a high school student. The test is currently a single-use product.
• There is a possibility that a women’s menstrual cycle may interfere with the results of the test. The
presence of blood currently has an unknown effect on the results of the assay due to limited
research.
• The assay and product design would need further testing and calibration prior to mass production.
This includes exploring extended shelf life of the product.
Figure 1: Results of the self-administered survey.
Figure 2: Digitally rendered 3D model of the prototype design.
How the Rapid Vaginitis Test Functions
Step 1
The sterile swab is removed from its packaging and
used as directed by swabbing the infected area. The
swab is then inserted into the solution chamber, and
stirred. The swab is screwed on and the test is left to
incubate at room temperature for 10 minutes to allow
the solution to react.
Screw on Swab
Holder
Sterile Swab
*Disclaimer: Models depicted in this diagram are for visual purposes only.
Figure 3: Flow chart and interal product diagram depicting the testing process.
Figure 4: Functional electronic circuit schematic to analyze and present the results of the assay.
Note: All diagrams and illustrations were created by the author.
Survey Results
• 36.5% of respondents who took the survey were unaware of what a yeast infection is, 25.0% of those
being female.
• 41.7% of students said that if they had a potential yeast infection, they were unsure or would not seek
professional treatment (see Figure 1).
• Embarrassment was the highest reported barrier at 52.8% followed by general discomfort with the
concept at 45.8% as seen in Figure 1.
• 59.7% of respondents would prefer to use an at-home test kit over going to the doctor (see Figure 1).
• 87.5% of respondents were interested in learning more about diagnosing, treating, and general
information about yeast infections.
Prototype Design
• An at-home rapid vaginitis test can be created by testing for the presence of bacteria.
• This can be done by testing for the presence of an enzyme called sialidase, which is commonly seen
with bacterial vaginosis.
• Research published in Nature found a biochemiluminescent sialidase assay that uses a substrate
including firefly luciferin that can accurately detect the presence of the enzyme [8]. In the presence of
sialidase, the substrate is cleaved to release luciferin, which is oxidized by firefly luciferase to generate
a light signal. This process is depicted in Figue 3. The intensity of the light signal can be used to
measure the concentration of sialidase.
• All reagents are present in a reagent bead and sample buffer, enabling a one-step assay. The one-step
assay enables a compact and easy-to-use product to be designed to create a rapid vaginitis test.
• In the clinical study conducted in the Nature article, the test tube was placed in a Helios 2000
luminometer to measure the light signal. Bacterial vaginosis is diagnosed when the signal is at or
greater than the cutoff value of 400,000 relative light units (RLU) [8].
• As a luminometer is an inaccessible piece of lab equipment, a cost-effective circuit can be designed to
measure the light signal intensity using a phototransistor, as seen in Figure 4. An experimental study
would need to be conducted to determine the new cut-off value, as RLUs are not convertible to lux or
lumen.
• The final rendition of the design includes a solution chamber and an electronics chamber separated by
transparent plastic to allow the phototransistor to detect light. The detailed design of the circuit that
would be used to analyze the results can be seen in Figure 4.
• The product would supply the user with the test, a sterile swab with a holder, and instructions.
• The test would function as follows: Using the sterile swab, pinch the holder and swab the suspected
infected area clockwise for 10 seconds. Unscrew the cap on the test and insert the swab into the
solution chamber, stirring before screwing the holder onto the test. Allow the test to incubate for 10
minutes after which the result will be displayed on the digital screen. If the test indicates the presence
of a bacterial infection, it is advised to seek medical assistance. This process can be seen in Figure 3.
Step 2 Step 3 • The estimated cost for a singular test is less than 10 dollars if mass-produced.
The assay detects the presence of bacteria by testing The light emissions are read by a phototransistor, a
for the enzyme Sialidase. Sialidase cleaves the device that is able to sense light levels and amplifies Protocol
substrate sialic acid – O –luciferin (SA-Firefly Luciferin) current. The phototransistor converts the light levels
to release luciferin, which is subsequently oxidized by
luciferase in the reaction to produce a detectable light
into a current which the bias resistor then turns into
voltage based on preset sensitivity. The resulting
• The one-step assay is created by combining two key elements. First, a lyophilized reagent bead is
created using thirty (30) microliters of a concentrated master mix solution.
signal. The light signal indicates the presence of voltage is then compared to the baseline value and
sialidase in the sample. results are displayed on the screen. If the results
match or are above the determined baseline the
screen reads “Bacterial Infection”. If the results are
• The master mix solution is created using the following ratios: 1% BSA, 3 mM ATP, 10 mM DTT, 1 mM
below the determined baseline the screen reads “Not
Bacterial Infection”.
coenzyme A, 1 mM substrate, and 4 micrograms/mL luciferase.
• Second, a sample buffer is created containing the following ratios: 50 mM HEPES (pH 7.0–7.2), 15 mM
SA- Firefly Luciferin
magnesium sulfate, 4 mM calcium chloride, 0.5% BSA, 0.5% Triton X-100, and 0.1% Proclin 950.
Sialidase
Bacterial
Firefly Luciferin Infection
• To perform the sialidase assay for a sample, a vaginal swab is suspended in 1 mL of sample buffer, and
OR two hundred fifty (250) microliters of the resulting sample are added to a reagent bead and incubated
Sialic Acid
Byproducts Luciferase at room temperature for 10 minutes.
Not Bacterial
Infection
Conclusions
Light
Firefly Oxyluciferin
Transparent Plastic
to Measure Light
Sialidase Detection
Assay Solution
Emission Phototransistor Bias Resistor Microcontroller Results Screen Battery This study supplies the necessary reasoning, ingredients, models, and instructions required to
manufacture an accessible and effective rapid vaginitis test kit.
Importantly, this study provides the methods and the design to create a mass-producible product to
the benefit of a broad demographic. The results of the self-conducted survey show 59.7% of respondents
would prefer to use an at-home test kit over going to the doctor. The estimated cost of less than 10
dollars makes the product accessible to low-income households or use in clinics. This product provides a
solution to those who cannot afford an immediate doctor’s visit and lab testing.
Given that 36.5% of students who took the survey were unaware of what a yeast infection is and
87.5% of respondents were interested in learning more about yeast infections, this study highlights the
need to create educational pamphlets and posters. This gives readers the tools to learn more about the
causes, treatments, and statistics surrounding vaginitis to protect their health. Education and an at-home
rapid testing product will reduce the stigma of vaginitis and promote wellbeing for all.
References
1. Centers for Disease Control and Prevention. (2021, October 12). Vaginal Candidiasis. Centers for Disease Control and Prevention. Retrieved December 1, 2021,
from https://www.cdc.gov/fungal/diseases/Candidiasis/genital/.
2. Cleveland Clinic. (2019, November 26). Vaginal yeast infections: Treatment, causes, prevention & symptoms. Cleveland Clinic. Retrieved December 1, 2021, from
https://my.clevelandclinic.org/health/diseases/5019-yeast-infections.
3. Frothingham, S. (2020, March 27). Untreated yeast infection: Why it should be diagnosed and treated. Healthline. Retrieved December 30, 2021, from
https://www.healthline.com/health/untreated-yeast-infection
4. Indeed Editorial Team. (2021, July 21). What are the 7 steps of the Engineering Design Process? Indeed Career Guide. Retrieved December 30, 2021, from
https://www.indeed.com/career-advice/career-development/design-process
5. Mayo Foundation for Medical Education and Research. (2021, March 17). Yeast infection (vaginal). Mayo Clinic. Retrieved December 29, 2021, from
https://www.mayoclinic.org/diseases-conditions/yeast-infection/symptoms-causes/syc-20378999
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https://pubmed.ncbi.nlm.nih.gov/29671516/
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