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1. Human papillomavirus (HPV) shedding in breast milk from African women living with HIV

Author

Jean-Pierre Molès, Marianne Peries, David Rutagwera, Philippe Van de Perre, James K Tumwine, Nathalie Boulle, Sophie Diaz, Chipepo Kankasa, Michel Segondy, Pathogénèse et contrôle des infections chroniques (PCCI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Universitaire de Montpellier (CHU Montpellier )-Université de Montpellier (UM), Laboratoire de Biologie cellulaire et Hormonale, Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Hôpital Arnaud de Villeneuve, Département d'oncologie médicale [CHU Montpellier], Centre Hospitalier Universitaire de Montpellier (CHU Montpellier ), Department of Paediatrics and Child Health, University of Zambia, Department of Paediatrics and Child Health [Lusaka, Zambia], and Makerere University [Kampala, Ouganda] (MAK)

Subjects
0301 basic medicine, Adult, medicine.medical_specialty, HPV, Breast milk, Hpv genotyping, Breastfeeding, Human immunodeficiency virus (HIV), Zambia, HIV Infections, medicine.disease_cause, 03 medical and health sciences, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Virology, Antiretroviral treatment, Medicine, Humans, Uganda, Human papillomavirus, Papillomaviridae, African women, Hpv types, Milk, Human, business.industry, Obstetrics, Papillomavirus Infections, virus diseases, HIV, 3. Good health, Virus Shedding, Hpv testing, 030104 developmental biology, Infectious Diseases, Breast Feeding, 030220 oncology & carcinogenesis, Africa, DNA, Viral, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, HIV-1, Female, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business
Abstract

Background Little is known about human papillomavirus (HPV) shedding in human breast milk. Objective To investigate HPV shedding in mature breast milk specimens collected from breastfeeding African women living with HIV-1 and not receiving antiretroviral treatment. Design 62 African women enrolled in the ANRS 12174 trial participated in this study. 79 lactoserum specimens obtained from right and/or left breasts from 42 Zambian women as well as lactosera and cell pellets from 40 milk samples collected from right and left breasts among 20 Ugandan women were tested for HPV using the INNO-LiPA HPV Genotyping Extra II assay. Results HPV DNA was detected in 9 (11.4%) lactoserum specimens collected from 8 (19.0%) Zambian women. Fourteen (17.5%) samples from 5 (25%) Ugandan women were positive for HPV detection. Differences in HPV type identification between the two breasts as well as between lactoserum and cell pellet were oberved. Overall, 13 (21.0%) of the 62 women included in this study had detectable HPV DNA in their breast milk, representing 11 HPV types, including high-risk, probable high-risk and low-risk types. Conclusion This study confirms that HPV can be frequently detected in breast milk in HIV-infected women. Further studies are needed to understand the way by which maternal milk can shed HPV.

Published
2018
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2. Multiple Hotspot Mutations Scanning by Single Droplet Digital PCR

Author

Charles Decraene, Olivier Lantz, Charlotte Proudhon, Audrey Vallée, Samia Melaabi, Adrien Saliou, Maud Milder, Amanda Bortolini Silveira, Alexandre Houy, Marc Michel, Marc-Henri Stern, François-Clément Bidard, Marc Ychou, Jean-Yves Pierga, Marc G. Denis, Anne Vincent-Salomon, Bernardo, Elizabeth, Laboratoire des biomarqueurs tumoraux circulants [Institut Curie, Paris] (SiRIC - Département de recherche translationnelle), Institut Curie [Paris]-Université Paris sciences et lettres (PSL), Compartimentation et dynamique cellulaires (CDC), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université Paris sciences et lettres (PSL), Département d'Oncologie Médicale [Institut Curie, Paris], Institut Curie [Paris], Clinical and Translational Research in Skin Cancer (CRCINA-ÉQUIPE 2), Centre de Recherche en Cancérologie et Immunologie Nantes-Angers (CRCINA), Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes), Département de Biochimie [CHU Nantes], Centre hospitalier universitaire de Nantes (CHU Nantes)-Hôpital Nord Laennec [CHU Nantes], Département de Biopathologie [Institut Curie, Paris], Génétique et Biologie du Développement, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Unité de génétique et biologie des cancers (U830), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre d'Investigation Clinique en Biotherapie des cancers (CIC 1428 , CBT 507 ), Institut Gustave Roussy (IGR)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunité et cancer (U932), Département d'oncologie médicale [CHU Montpellier], Centre Hospitalier Universitaire de Montpellier (CHU Montpellier ), Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Université Paris Descartes - Paris 5 (UPD5), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), and Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, Biopsy, Clinical Biochemistry, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, medicine.disease_cause, Polymerase Chain Reaction, Circulating Tumor DNA, 03 medical and health sciences, Exon, 0302 clinical medicine, [SDV.CAN] Life Sciences [q-bio]/Cancer, Limit of Detection, Neoplasms, medicine, Humans, Digital polymerase chain reaction, Liquid biopsy, Lung cancer, Gene, medicine.diagnostic_test, Biochemistry (medical), Liquid Biopsy, medicine.disease, Molecular biology, 3. Good health, ErbB Receptors, 030104 developmental biology, Genes, ras, 030220 oncology & carcinogenesis, Molecular Probes, Mutation, Cancer research, KRAS, Molecular probe
Abstract

BACKGROUND Progress in the liquid biopsy field, combined with the development of droplet digital PCR (ddPCR), has enabled noninvasive monitoring of mutations with high detection accuracy. However, current assays detect a restricted number of mutations per reaction. ddPCR is a recognized method for detecting alterations previously characterized in tumor tissues, but its use as a discovery tool when the mutation is unknown a priori remains limited. METHODS We established 2 ddPCR assays detecting all genomic alterations within KRAS exon 2 and EGFR exon 19 mutation hotspots, which are of clinical importance in colorectal and lung cancer, with use of a unique pair of TaqMan® oligoprobes. The KRAS assay scanned for the 7 most common mutations in codons 12/13 but also all other mutations found in that region. The EGFR assay screened for all in-frame deletions of exon 19, which are frequent EGFR-activating events. RESULTS The KRAS and EGFR assays were highly specific and both reached a limit of detection of CONCLUSIONS This method presents the advantage of detecting a higher number of mutations with single-reaction ddPCRs while consuming a minimum of patient sample. This is particularly useful in the context of liquid biopsy because the amount of circulating tumor DNA is often low. This method should be useful as a discovery tool when the tumor tissue is unavailable or to monitor disease during therapy.

Published
2017
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