Effects of LiveSpo X-secret in Supporting Treatment of Sexually Transmitted Diseases
Condition: Sexually Transmitted Diseases
MeSH terms: Sexually Transmitted Diseases
Study type: Interventional

Summary: 
      Sexually Transmitted Infections (STIs) are common gynecological infections worldwide,
      especially in tropical and underdeveloped countries, primarily affecting the reproductive age
      group. Microbial pathogens causing STIs include bacteria, viruses, fungi, and protozoa. In
      Vietnam, STIs are a significant public health concern due to their high incidence and
      tendency to have mild or no symptoms, making them easy to overlook. While antibiotics or
      antifungal drugs are typically used to treat STIs caused by bacterial or fungal agents such
      as Chlamydia, Gonorrhea, and Candida, no specific treatment exists for most viral STIs. Thus,
      vaccination and symptomatic treatment are the primary methods of prevention. The use of
      broad-spectrum antibiotics can result in side effects such as loss of appetite, fatigue, and
      sexual problems, while repeated treatments can lead to the emergence of antibiotic
      resistance, posing a significant challenge for STI treatment worldwide. However, the efficacy
      of oral or suppository-based probiotics is limited by their slow onset and sensitivity to
      acidic and temperature conditions. Fortunately, Bacillus strains of probiotics have
      demonstrated exceptional stability in both acidic and temperature environments, making them
      particularly well-suited for use as vaginal spray probiotics against STIs. Here,
      investigators propose that vaginal-spraying probiotics containing Bacillus strains could be a
      safe and effective symptomatic treatment for STIs caused by viral, fungi, or bacterial
      pathogens, providing a promising alternative to traditional antibiotic therapy.

      The aim of the study about to evaluate the effectiveness of vaginal-spraying probiotics
      containing spores of three strains of Bacillus subtilis, Bacillus clausii, and Bacillus
      coagulans in preventing and supporting the treatment of vaginal infections.

      Study Population: sample size is 100. The study is being carried out at Bac Ninh Center for
      Disease Control.

      Description of Study Intervention: total of 100 eligible patients are divided randomly into 2
      groups (n = 50/group): Patients in the Control group received routine treatment and 3
      times/day 0.9% NaCl physiological saline while the patients in the experimental group were
      treated with LiveSpo X-secret 3 times/day in addition to the same standard of care treatment.
      The standard treatment regimen is 7 days and is maintained for 28 days.

      Study duration: 36 months
    

Detailed description: 
      Sexually Transmitted Infections (STIs) are common gynecological infections worldwide,
      especially in tropical and underdeveloped countries where up to three-quarters of the
      population has STIs, mainly in the reproductive age. The infection rate in regions and
      countries is different, the infection rate in developing countries is 20-fold higher than in
      developed countries. Southeast Asia alone has about 150.5 million people with these diseases.
      Vietnam is located in the tropics, and hot and humid climates are favorable factors for
      microbial pathogens to develop. According to reports from the dermatology units of the
      provinces, every year about 300,000 people are infected with sexually transmitted
      diseases...There are many microbial pathogens causing infections including bacteria, viruses,
      fungi, and protozoa...The patient can be infected with multiple pathogens at the same time.
      This is a matter of great concern in public health care because STIs often have mild or no
      symptoms, so they are easily overlooked. While antibiotics or antifungal drugs are typically
      used to treat STIs caused by bacterial or fungal agents like Chlamydia, Gonorrhea, and
      Candida, no specific treatment exists for most viral STIs. However, the use of broad-spectrum
      antibiotics can result in side effects like loss of appetite, fatigue, and sexual problems,
      while repeated treatments can lead to the emergence of antibiotic resistance, posing a
      significant challenge for STI treatment worldwide. Probiotics are microorganisms that are
      beneficial to our health when consumed in adequate amounts, playing a vital role in
      maintaining a healthy balance of bacteria in our body, especially in the gut and vaginal
      areas. Studies have found that probiotics can help reduce the incidence and severity of
      vaginal infections, including bacterial vaginosis and yeast infections. Currently, oral or
      vaginal suppository probiotics containing strains of Bifidobacterium or Lactobacillus are
      being studied as a potential complementary therapy for treating vaginal infections. However,
      these strains have low survivability and are sensitive to light, moisture, and pH levels,
      making preservation challenging. Thus, their effectiveness depends on the specific strains
      used and the dose required to achieve colonization in the vaginal tract. Bacillus is a
      well-studied probiotic strain with over 350,000 research publications worldwide, and its
      safety has been extensively investigated. Bacillus subtilis, B. clausii, and B. coagulans are
      all aerobic or facultative anaerobic bacteria capable of spore formation and can tolerate
      harsh conditions such as nutrient deprivation, anoxia, drought, and even antibiotics. Here,
      investigators propose that vaginal-spraying probiotics containing Bacillus strains could be a
      safe and effective symptomatic treatment for STIs caused by viral, fungi, or bacterial
      pathogens, providing a promising alternative to traditional antibiotic therapy.

      The objective was to investigate the symptomatic treatment effects of the probiotic product
      LiveSpo X-secret as a liquid-suspension form containing Bacillus spores of safe B. subtilis
      ANA4, B. clausii ANA39, B. coagulans ANA 40 strains, in women having sexually transmitted
      infection and investigators evaluation of improved efficacy and reduced clinical symptoms
      treatment time and measurement of changes in pathogen load in the vaginal samples before
      treatment (day 0) and after 3, 7, 14 and 28 days using LiveSpo X-secret.

      Methods: A randomized, blind, and controlled clinical trial is conducted. After informed
      consent, patients will be screened for common nine STI pathogens in Vietnam including
      Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis,
      Mycoplasma genitalium, Human alphaherpesvirus type 1 and 2, Candida albicans, Trichomonas
      vaginalis. Totally 100 patients positive with at least one of nine STIs will be randomized
      into 2 groups (n = 50/group): the control group (named "Control" group) uses 0.9% NaCl
      physiological saline and an experimental group (named the "X-secret" group) use the
      probiotics LiveSpo X-secret. The patient is given a coded spray in the form of a blind sample
      to ensure the objectivity of the study. The clinical follow-up will be at the time point of
      0, 3, 7, 14, and 28 days, vaginal samples will be collected at days 0, 3, 7, 14, and day 28
      to evaluate potential reductions in pathogens load and change the microbiome in vaginal
      flora.

      Real-time PCR for detection of microorganism in vaginal samples: semi-quantitative assays for
      measuring changes in pathogens load is conducted by the real-time PCR assay which has been
      optimized and developed by the research team (Bui et al., 2023). Detection of useful Bacillus
      strains belonging to B. subtilis, B. clausii, and B. coagulans is conducted by real-time PCR
      SYBR Green assay that is routinely runs also conducted at Spobiotic Research Center, Hanoi,
      Vietnam

      Analysis of vaginal microbiome 16S rRNA using next-generation sequencing (NGS) at Macrogene
      (Seoul, Korea) on the Illumina MiSeq (Illumina, San Diego, CA, USA) using a 2 × 250 bp run
      configuration.

      During treatment, patients are monitored for typical clinical symptoms of gynecological
      infections, including itching, odor, vaginal color, and pH from vaginal samples of Control
      and X-secret groups at the time points of day 3, 7, 14, and day 28. The patients' health
      conditions are observed by doctors and nurses, and their pieces of information are filled
      into medical records at the patient's follow-up visits or phone interviews.

      Data collection and statistical analysis: individual medical records are collected, and the
      patient's information is then gathered and systematized in a data set. The safety and
      efficacy of LiveSpo X-secret are evaluated and compared to 0.9% NaCl physiological saline
      based on the following clinical and sub-clinical criteria obtained in X-secret and Control
      groups: (i) the symptomatic-relieving day and level of symptom relief; (ii) the reduction
      levels (2^△Ct) of pathogens load △Ct for target genes is calculated as Ct (threshold cycle)
      at day 3/7/14/28 - Ct at day 0 while Ct of internal control is adjusted to be equal among all
      samples; (iii) change the microbiome in vaginal flora.

      The tabular analysis is performed on dichotomous variables using the χ2 test or Fisher's
      exact test when the expected value of any cell is below five. Continuous variables are
      compared using either the Wilcoxon test, t-test, or the Mann-Whitney test when data are not
      normally distributed. Statistical and graphical analyses are performed on GraphPad Prism
      v8.4.3 software (GraphPad Software, CA, USA). The significance level of all analyses is set
      at p < 0.05. P-values.

      Expected outcomes: (i) LiveSpo X-secret alleviates influenza-infection symptoms about 30%
      more effectively, as indicated by 90% of patients using LiveSpo X-secret (X-secret group) are
      symptom-free at day 3-28 of intervention depending on symptoms, compared to 60% of patients
      in Control group; (ii) Patients in X-secret group has more significant reductions in
      pathogens load (>10 fold) than patients in Control group at the time point of day 3, 7, 14,
      28 of intervention.
    

Intervention name(s): LiveSpo X-secret; 0.9% NaCl physiological saline
Intervention type(s): Combination Product; Drug
Intervention description(s): In Vietnam, LiveSpo X-secret is manufactured as a Class-A medical device product (Product declaration No.220003286/PCBA-HN) under manufacturing standards approved by Hanoi Health Department, Ministry of Health, Vietnam (Certificate No YT117-19) and ISO 13485:2016.; Vaginal-spraying 0.9% NaCl physiological saline is prepared by extracting 10 mL from 0.9% NaCl intravenous infusion 500 mL PP bottle (B.Braun, Germany, product declaration No. VD-32732-19), and then pouring it into the same opaque metallic spraying 15 mL-bottle that is used for LiveSpo X-secret.
Other intervention name(s): Registration number: No.220003286/PCBA-HN; Registration number: VD-32723-19
Allocation: Randomized
Intervention model: Parallel Assignment
Intervention model description: Parallel Assignment Blind, randomized, and controlled clinical trial
Masking: Triple (Participant, Care Provider, Investigator)
Primary purpose: Treatment
Keywords: Sexually Transmitted Diseases (STDs); Sexually Transmitted Infections (STIs); Bacterial vaginosis (BV); Vaginal-spraying probiotics; Bacillus spores; Pathogen load; Fungi; Virus; Vietnamese women