Maternal- Fetal Infection

Study Overview
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Brief Summary
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The purpose of the protocol is to validate a novel point of care multiplex system to detect and characterize microorganisms responsible for neonatal sepsis, as well as biomarkers of infection, from a simple vaginal sample, in order to improve the prevention of perinatal bacterial infections.

Detailed Description
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Early-onset neonatal sepsis (EOS) is a major global public health challenge. Prevention during pregnancy and delivery, early diagnosis and treatment of perinatal infections are essential to avoid EOS. Risk factors for include prematurity, maternal Group B streptococcus (GBS) colonization, premature rupture of membranes (PROM), and chorioamnionitis. Screening and intrapartum antimicrobial prophylaxis administered to GBS-colonized women has reduced early onset GBS infections. However, other pathogens are frequently involved in EOS following preterm PROM and preterm birth (PTB), such as Gram-negative bacteria and Staphylococci, which are not covered by penicillin prophylaxis. The prevalence of neonatal infection arising from antibiotic-resistant bacteria is increasing, thus the challenge is to eliminate the widespread unnecessary use of broad-spectrum antibiotics to treat non-infected infants, while recognizing when antibiotics are truly needed. Rapid diagnostic test(s) to detect and quantify specifically pathogens in vaginal samples, could be a major breakthrough. Several rT- PCR ( reverse Transcriptase Polymerase Chain Reaction) tests are on the market, however so far no test is able to detect, quantify and characterize in terms of antibiotic resistance and virulence genes, a range of pathogens. A novel multiplex platform, using microfluidics technology, is under development by Elvesys, Inc in France. This platform will be able to offer results within 15 minutes on-site. In addition, the study of the vaginal microbiome may identify signatures associated with a risk of maternal-fetal infection, particularly in case of PROM or PTB. Advanced sequencing technology and metagenomics will be used to characterize these signatures, and may lead to further markers to be included in the point-of-care test. Finally, biomarkers of inflammation will be detected, including IL-6 (Interleukin). In this study, the InSPIRe platform will be compared in the laboratory to conventional microbiological and immunological detection. Four groups of pregnant women will be recruited in prospective cohorts : uneventful pregnancies, term PROM, preterm labor and preterm PROM. The purpose of the InSPIRe project is to improve the prevention of perinatal bacterial infections, with the novel Elvesys point of care system to rapidly detect and characterize microorganisms responsible for neonatal sepsis from a single vaginal sample.

Official Title
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Innovative Strategies for Perinatal Infectious Risk Reduction

Conditions
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Neonatal Infection

Intervention / Treatment
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* Biological: Bacteriological analyses on clinical samples performed with swabs


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Pregnant woman, Gestational age over 22 SA, Patient agreeing to sign informed consent, Patient aged at least 18 years old, Patient with health insurance, Singleton, twin or multiple pregnancy. Exclusion Criteria: Fetal death or non-viable fetus, maternal age under 18, Patient unable to express her consent, Patient under guardianship.

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Screening
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: There are 4 pre-specified groups
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Woman at low risk of infection<br>Women with systematic vaginal sample for detection of GBS will be included. | Biological: Bacteriological analyses on clinical samples performed with swabs<br>* Bacteriological analyses will be performed to assess the InSPIRe kit<br>|
| Experimental: Woman with high risk of infection > 37 SA<br>Women with premature rupture of membranes (> 12 hours before labor) but > 37 SA will be included. | Biological: Bacteriological analyses on clinical samples performed with swabs<br>* Bacteriological analyses will be performed to assess the InSPIRe kit<br>|
| Experimental: Women with premature rupture of membranes (<37SA)<br>Woman with high risk of infection <37SA | Biological: Bacteriological analyses on clinical samples performed with swabs<br>* Bacteriological analyses will be performed to assess the InSPIRe kit<br>|
| Experimental: Women with premature delivery or premature delivery threat<br>Woman with high risk of infection <37SA and Women with premature delivery or premature delivery threat | Biological: Bacteriological analyses on clinical samples performed with swabs<br>* Bacteriological analyses will be performed to assess the InSPIRe kit<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Presence of Streptococcus B |  | Day 0 | 
| Presence of Streptococcus B |  | until 20 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Maternal fetal infection | Infection is proved if at least a sample, generally sterile, is positive to a germ in association with a positive clinical, biological or radiologic sign of infection such as C-reactive protein or chest radiography. | until 20 weeks + 3 days | 
| A positive bacteriological result in the vaginal sample | A positive result is defined as the presence of one of the subsequent germ in the bacteriological culture: Escherichia coli (E. coli) Streptococcus pneumoniae, Group A Streptococcus Haemophilus ssp (influenzae, parainfluenzae) Staphylococcus aureus Streptococcus milleri group Enterococcus faecalis Other Gram-negative bacilli type enterobacteria (Klebsiella pneumonia, Proteus mirabilis) Anaerobics (Prevotella sp, bacteroid fragilis) | Day 0 | 
| A positive bacteriological result in the vaginal sample | A positive result is defined as the presence of one of the subsequent germ in the bacteriological culture: Escherichia coli (E. coli) Streptococcus pneumoniae, Group A Streptococcus Haemophilus ssp (influenzae, parainfluenzae) Staphylococcus aureus Streptococcus milleri group Enterococcus faecalis Other Gram-negative bacilli type enterobacteria (Klebsiella pneumonia, Proteus mirabilis) Anaerobics (Prevotella sp, bacteroid fragilis) | until 20 weeks | 
| Vaginal dysmorphism | Defined by lactobacillus's decrease or loss in association with the spread of anaerobic flora. | day 0 | 
| Vaginal dysmorphism | Defined by lactobacillus's decrease or loss in association with the spread of anaerobic flora. | until 20 weeks | 
| Antibiotic resistance | Highlighted by a resistant profile in bacteriological culture for various classes of antibiotics such as β-lactamines, macrolides or aminoamides. | Day 0 | 
| Antibiotic resistance | Highlighted by a resistant profile in bacteriological culture for various classes of antibiotics such as β-lactamines, macrolides or aminoamides. | until 20 weeks | 
| Highlighting specific virulence markers | Like GBS clone CC17, Enterobacteriaceae that produce capsular antigen type K1. By usual molecular techniques. | Day 0 | 
| Highlighting specific virulence markers | Like GBS clone CC17, Enterobacteriaceae that produce capsular antigen type K1. By usual molecular techniques. | until 20 weeks | 
| Maternal local biomarkers definition | Presence or lack of RNA sequences and/or human specific protein detected by RT-PCR or ELISA-PCR depending on the presence or lack of a proved or suspected maternal fetal infection. | Day 0 | 
| Maternal local biomarkers definition | Presence or lack of RNA sequences and/or human specific protein detected by RT-PCR or ELISA-PCR depending on the presence or lack of a proved or suspected maternal fetal infection. | until 20 weeks | 
| Bacteriological signature definition | Presence or lack of specific bacteriological sequences detected by global sequencing and metagenomics analyses | Day 0 | 
| Bacteriological signature definition | Presence or lack of specific bacteriological sequences detected by global sequencing and metagenomics analyses | until 20 weeks | 
| Chorioamnionitis | Clinical or paraclinical factors associated with risk of chorioamnionitis or maternal fetal infection such as prematurity, clinical signs (maternal fever, fetal tachycardia, increase in C-reactive protein, hyperleukocytosis, pus-like amniotic fluid) , oligohydramnios (defined by the greatest cistern < 25 mm); increase in pro-calcitonin in umbilical cord ok histological signs of placental inflammation. Clinical chorioamnionitis is defined as a maternal fever> 39° (in one shot) with no other cause or >38°(confirmed) in association with abnormal fetal heartbeat (>160/min exceeding 10 min), maternal hyperleukocytosis (>15000/mm3 without corticotherapy) or pus-like amniotic fluid. Histological chorioamnionitis is defined by the presence of neutrophil polynuclears in amnion or chorion in association with the presence of neutrophil polynuclears in umbilical cord blood vessels. | Day 0 | 
| Chorioamnionitis | Clinical or paraclinical factors associated with risk of chorioamnionitis or maternal fetal infection such as prematurity, clinical signs (maternal fever, fetal tachycardia, increase in C-reactive protein, hyperleukocytosis, pus-like amniotic fluid) , oligohydramnios (defined by the greatest cistern < 25 mm); increase in pro-calcitonin in umbilical cord ok histological signs of placental inflammation. Clinical chorioamnionitis is defined as a maternal fever> 39° (in one shot) with no other cause or >38°(confirmed) in association with abnormal fetal heartbeat (>160/min exceeding 10 min), maternal hyperleukocytosis (>15000/mm3 without corticotherapy) or pus-like amniotic fluid. Histological chorioamnionitis is defined by the presence of neutrophil polynuclears in amnion or chorion in association with the presence of neutrophil polynuclears in umbilical cord blood vessels. | until 20 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Neonatal infections, maternal-fetal infection, group B streptococcus, vaginal sample, antibiotic prophylaxis, bacterial resistance