Laboratory development of a simple stool sample processing method diagnosis of pediatric tuberculosis using Xpert Ultra
Manon Lounnas
(1, 2)
,
Abibatou Diack
(1, 2)
,
Mark P. Nicol
(3, 4)
,
Sara Eyangoh
(5)
,
Eric Wobudeya
(6)
,
Olivier Marcy
(7, 8)
,
Sylvain Godreuil
,
Maryline Bonnet
(9, 10)
1
MIVEGEC -
Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle
2 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]
3 UWA - The University of Western Australia
4 University of Cape Town
5 Centre Pasteur du Cameroun
6 MAK - Makerere University [Kampala, Ouganda]
7 BPH - Bordeaux population health
8 GHiGS - Global Health in the Global South
9 TransVIHMI - Recherches Translationnelles sur le VIH et les maladies infectieuses endémiques et émergentes
10 Epicentre [Paris] [Médecins Sans Frontières]
2 CHRU Montpellier - Centre Hospitalier Régional Universitaire [Montpellier]
3 UWA - The University of Western Australia
4 University of Cape Town
5 Centre Pasteur du Cameroun
6 MAK - Makerere University [Kampala, Ouganda]
7 BPH - Bordeaux population health
8 GHiGS - Global Health in the Global South
9 TransVIHMI - Recherches Translationnelles sur le VIH et les maladies infectieuses endémiques et émergentes
10 Epicentre [Paris] [Médecins Sans Frontières]
Manon Lounnas
- Fonction : Auteur
- PersonId : 1358375
- IdHAL : manon-lounnas
- ORCID : 0000-0001-8789-0517
Sara Eyangoh
- Fonction : Auteur
- PersonId : 1182761
- ORCID : 0000-0002-0557-3815
Olivier Marcy
- Fonction : Auteur
- PersonId : 1218137
- ORCID : 0000-0003-3350-112X
Sylvain Godreuil
- Fonction : Auteur
- PersonId : 1147481
- ORCID : 0000-0003-3436-0082
- IdRef : 069435847
Maryline Bonnet
- Fonction : Auteur
- PersonId : 1237206
- ORCID : 0000-0002-1071-7393
Résumé
Stool samples are alternatives to respiratory samples for bacteriological confirmation of childhood tuberculosis but require intensive laboratory processing before molecular testing to remove PCR inhibitors and debris. We aimed to develop a centrifuge-free processing method for use in resource-limited settings based on a sucrose-flotation method that showed good sensitivity for childhood tuberculosis diagnosis.In an in vitro study using Xpert MTB/RIF Ultra on stool samples spiked with defined bacterial concentrations of Mycobacterium tuberculosis (MTB), we compared different simplification parameters to the reference sucrose-flotation method. Best methods were selected based on the rate of invalid/error results and on sensitivity, compared to the reference method on stools spiked at 103 colony forming units (CFU)/g MTB. For final selection, we tested the best parameter combinations at 102 CFU/g. Out of 13 different parameter combinations, three were tested at 102 CFU/g. The best combination used 0.5 g stool, manual shaking, no filtration, 30-min sedimentation, and a 1:3.6 dilution ratio. This method gave 10% invalid/error results and a sensitivity of 70% vs 63% at 103 CFU/g and 53% vs 58% at 102 CFU/g compared to the reference method.This pre-clinical study was able to develop a centrifuge-free processing method to facilitate stool Xpert Ultra testing.
Format du dépôt | Fichier |
---|---|
Type de dépôt | Article dans une revue |
Titre |
en
Laboratory development of a simple stool sample processing method diagnosis of pediatric tuberculosis using Xpert Ultra
|
Résumé |
en
Stool samples are alternatives to respiratory samples for bacteriological confirmation of childhood tuberculosis but require intensive laboratory processing before molecular testing to remove PCR inhibitors and debris. We aimed to develop a centrifuge-free processing method for use in resource-limited settings based on a sucrose-flotation method that showed good sensitivity for childhood tuberculosis diagnosis.In an in vitro study using Xpert MTB/RIF Ultra on stool samples spiked with defined bacterial concentrations of Mycobacterium tuberculosis (MTB), we compared different simplification parameters to the reference sucrose-flotation method. Best methods were selected based on the rate of invalid/error results and on sensitivity, compared to the reference method on stools spiked at 103 colony forming units (CFU)/g MTB. For final selection, we tested the best parameter combinations at 102 CFU/g. Out of 13 different parameter combinations, three were tested at 102 CFU/g. The best combination used 0.5 g stool, manual shaking, no filtration, 30-min sedimentation, and a 1:3.6 dilution ratio. This method gave 10% invalid/error results and a sensitivity of 70% vs 63% at 103 CFU/g and 53% vs 58% at 102 CFU/g compared to the reference method.This pre-clinical study was able to develop a centrifuge-free processing method to facilitate stool Xpert Ultra testing.
|
Sous-Titre |
fr
Tuberculosis (Edinb)
|
Auteur(s) |
Manon Lounnas
1, 2
, Abibatou Diack
1, 2
, Mark P. Nicol
3, 4
, Sara Eyangoh
5
, Eric Wobudeya
6
, Olivier Marcy
7, 8
, Sylvain Godreuil
, Maryline Bonnet
9, 10
1
MIVEGEC -
Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle
( 166573 )
- 911 Avenue Agropolis BP 64501 34394 Montpellier cedex 5
- France
2
CHRU Montpellier -
Centre Hospitalier Régional Universitaire [Montpellier]
( 258773 )
- 191 avenue du doyen Gaston Giraud 34295 Montpellier
- France
3
UWA -
The University of Western Australia
( 4177 )
- 35 Stirling Highway
Perth WA 6009 Australia
- Australie
4
University of Cape Town
( 303907 )
- Private Bag X3, Rondebosch 7701, South Africa
- Afrique du Sud
5
Centre Pasteur du Cameroun
( 55917 )
- B.P. 1274 Yaoundé
- Cameroun
6
MAK -
Makerere University [Kampala, Ouganda]
( 328028 )
- P.O. Box 7062 - Kampala
- Ouganda
7
BPH -
Bordeaux population health
( 476572 )
- Université de Bordeaux - Case 11 - 146 rue Léo Saignat 33076 Bordeaux cedex
- France
8
GHiGS -
Global Health in the Global South
( 1162866 )
- Centre de recherche INSERM U1219 - 146 rue Léo-Saignat - 33076 BORDEAUX cedex 200
- France
9
TransVIHMI -
Recherches Translationnelles sur le VIH et les maladies infectieuses endémiques et émergentes
( 160874 )
- Centre IRD France Sud 911, avenue Agropolis BP 64501 F-34394 Montpellier cedex 5
- France
10
Epicentre [Paris] [Médecins Sans Frontières]
( 335654 )
- 55 Rue Crozatier, 75012 Paris
- France
|
Date de publication |
2020-12
|
Licence |
Paternité - Pas d'utilisation commerciale
|
Langue du document |
Anglais
|
Nom de la revue |
|
Vulgarisation |
Non
|
Comité de lecture |
Oui
|
Audience |
Internationale
|
Volume |
125
|
Page/Identifiant |
102002
|
Domaine(s) |
|
Mots-clés (Mesh) |
|
Mots-clés |
en
Childhood tuberculosis, Diagnosis, Stool, Xpert MTB/Rif ultra
fr
IDLIC
|
DOI | 10.1016/j.tube.2020.102002 |
PII | S1472-9792(20)30169-4 |
Pubmed Id | 33049437 |
UT key WOS | 000594211900010 |
Origine :
Fichiers produits par l'(les) auteur(s)
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