1.Method validation study for SARS-CoV-2 viral RNA detection in cervical, rectal, amniotic fluid, placental, umbilical cord blood, and breastmilk specimens in a cohort of unvaccinated women in Manila, Philippines
Erlidia F. Llamas-Clark ; Mayan U. Lumandas ; Daniel C. Villarico ; Amalea Dulcene D. Nicolasora ; Maria Stephanie Fay S. Cagayan ; Emmanuel S. Baja ; Maria Esterlita T. Villanueva-Uy ; Paulyn Jean B. Rosell-Ubial ; Francisco M. Heralde III
Acta Medica Philippina 2024;58(15):32-38
OBJECTIVES
To validate a method in detecting SARS-CoV-2 via RT-qPCR in pregnant and non-pregnant samples other than nasopharyngeal swabs and/or oropharyngeal swabs such as cervical, rectal, amniotic fluid, placental, umbilical cord blood, and breastmilk.
METHODSWe performed a validation experiment using MGI easy extraction kits and BGI PCR kits on non-conventional specimens, including cervical, rectal, amniotic fluid, placental, umbilical cord blood, and breastmilk to detect and confirm the presence of SARS-CoV-2. In addition, we tested the validated method on 572 purposively sampled field-collected non-conventional specimens from a cohort of 109 unvaccinated pregnant and 47 unvaccinated non-pregnant women to assess which candidate non-conventional maternal- and fetal-associated specimens may contribute to maternal-fetal viral vertical transmission.
RESULTSPositive detection of SARS-CoV-2 viral RNA in non-conventional specimens was demonstrated and verified. Of the 572 non-conventional samples tested, 1.8% (10/572) were positively validated by RT-qPCR for SARS-CoV-2 in the maternal-associated specimens particularly the rectal (5), placental (1), and cervical (4) swabs among six pregnant and four non-pregnant individuals. In contrast, no SARS-CoV-2 viral RNA was detected in fetal-associated specimens.
CONCLUSIONThe results of the validation study may serve as an additional diagnostic screening layer to support maternal-child care. Furthermore, viral detection in these non-conventional maternal specimens may also be utilized to provide guidance in the clinical management of neonates, and pregnant women during delivery.
Philippines ; Sars-cov-2 ; Pregnant Women ; Umbilical Cord ; Amniotic Fluid ; Polymerase Chain Reaction ; Placenta