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Newborn Screening for Maple Syrup Urine Disease (MSUD) using a totally automated system.

Authors:

Abstract

Introduction: In the last years the organization called Rede DXB, an MSUD workgroup of specialized professionals in Brazil, has produced actions for development of research and diagnosis of MSUD. MSUD is not mandated by the Brazilian Newborn Screening Program but published data reveals an incidence 5 times higher than expected. The Brazilian industry, looking to meet the requests and needs of the Newborn Screening Program, has developed a totally automated system for the detection of MSUD. Materials and methods : This study was developed using NeoLISA MSUD kit, routine newborn screening samples from different labs in Brazil, positive confirmed samples, internal controls and external quality control materials. The assay is enzymatic colorimetric and the procedure occurs in a totally automated system. The system takes 1 hour to run about 500 samples but can perform almost 800 samples per routine, or 3000 samples per day per equipment. Results : The results demonstrated the reproducibility of samples and controls used in the study including the external quality control materials. All positive samples were correctly classified as positives by NeoLISA MSUD. These samples were previously confirmed by HPLC method associated with clinical diagnosis. Conclusions : The enzymatic colorimetric method (NeoLISA MSUD Kit) associated with totally automated system (Nimbus NeoLISA) is an excellent alternative to meet the needs of the Brazilian Newborn Screening Program. The use of tMS/MS for MSUD and other diseases has been studied by many labs in Brazil. High cost associated with the equipment and accessories, human resources, infrastructure of the labs, and technical support are considered problematic and usually associated with the negative perspective to introduce MSUD testing. The use of NeoLISA MSUD meets the perspective proposed by the studies of Rede DXB with development of newborn screening program recognizing the professional’s efforts to prevent deleterious consequences in non-treated MSUD patients.
Newborn Screening for Maple Syrup Urine Disease (MSUD)
Using a Totally Automated System.
Claudio Sampaio Filho (Jr)1, Fabiola Frias1, Raquel Veturiano1,
Vanessa Silva1, Bruna Gallo1, Fabio Bernardes1, Jonatas Boalento1,
Anakele Massi1, Josias Lira1,
1-INTERCIENTIFICA, S.J.Campos - Brasil
Introduction:
Few years ago the organization, Rede DXB, a Brazilian MSUD Organization composed of a
workgroup of specialized professionals, took action to develop research and diagnosis of MSUD.
MSUD is not mandatory by the Brazilian Newborn Screening Program even though published data
reveals an incidence five times higher than expected. The Brazilian industry, aiming to meet the
requests and needs of the Newborn Screening Program, developed a totally automated system for
MSUD newborn screening . This system uses the enzymatic colorimetric method (NeoLISA®
MSUD kit) to quantify Leucine/Isoleucine in dot blood spots (DBS) collected in newborn screening
programs.
The main objective of this study is to demonstrate the availability of the assay in a totally
automated process, which is very similar to the protocols used for Phenylketonuria (PKU),
Galactosemia (GAL) and other enzymatic colorimetric methods commonly used in the routine. The
study shows evidences that the industry is ready to offer the best solution for newborn screening of
MSUD for Brazilian labs and other international labs interested to invest in the prevention of the
disease without the high costs associated with MS/MS.
Nº PO
Materials and Methods:
This study was developed using NeoLISA®MSUD kit, routine newborn screening samples from
different labs in Brazil, confirmed positive samples, internal controls and external quality control
materials. The enzymatic colorimetric assay is carried out in a totally automated system which
takes only one hour to run about 500 samples and can proceed with almost 800 samples per
routine, or 3000 samples per day per equipment.
Fig. 1 Nimbus NeoLISA® a Totally Automated System for newborn screening of MSUD offered by
INTERCIENTIFICA. The equipment is the same used for PKU, GAL, G6PD and BIO screening assays and can run
the assays simultaneously according to the lab needs. The equipment is capable of running 8 plates at same
time. The equipment is ready for use and does not require specific space, temperature control, or any specific
additional power source and needs as MS/MS. The NeoLISA® kits for MSUD used in this study as well as the
other enzymatic colorimetric assays used NeoLISA® / PKU / GAL / G6PD / BIO are produced by
INTERCIENTIFICA in Brazil under GMP Certificate by ANVISA. All these products are registered in ANVISA as in
vitro diagnostic products. NeoLISA® products are commercialized in Brazilian market for more than 20 years.
Newborn Screening for Maple Syrup Urine Disease (MSUD)
Using a Totally Automated System.
Claudio Sampaio Filho (Jr)1, Fabiola Frias1, Raquel Veturiano1,
Vanessa Silva1, Bruna Gallo1, Fabio Bernardes1, Jonatas Boalento1,
Anakele Massi1, Josias Lira1,
1-INTERCIENTIFICA, S.J.Campos - Brasil
Nº PO
Results:
The results demonstrated the reproducibility of samples and controls used in the study including
the external quality control materials.
Table 1. We produced controls at four different enrichment levels from 2 to 8 mg/dL. These 4 controls were used
for 20 runs, with 3 different lots of product NeoLISA®MSUD, and a total of 240 results. The external quality
control materials used from CDC (Center for Diseases,Control and Prevention) with 4 different levels. The
results of the lot 1425,1426,1427, 1428, average in 5 runs in duplicate, respectively, 1.9/3.4/5.1/7.0 mg/dL.
All positive samples (n=5) were correctly classified as positive by NeoLISA®MSUD. These
samples have been previously confirmed by HPLC method associated with clinical diagnosis. The
normal samples added to this study obtained an average of 1.95 mg/dL (n=1.932).
The Nimbus NeoLISA®is a totally automated system developed to run routines of 8 microplates
or around 720 samples in 3 hours and 15 minutes. The NeoLISA®MSUD kit uses the same assay
protocol of the other enzymatic colorimetric methods offered by Intercientifica (PKU/GAL/G6PD/
BIO) and can be integrated in the same equipment simultaneously.
Fig 2. Typical ISO/LEU standard curve using Nimbus NeoLISA®,results units mg/dL, one standard curve for the
entire routine.
Fig 3. Totally automated Process time for Nimbus NeoLISA®with 8 microplates in a routine. The different colors
demonstrate the different procedures in time periods.
ISSO/LEU (mg/dL) INTRA-ASSAY INTER-ASSAY
A 2.0 3.1 3.8
B 4.0 4.7 5.1
C 6.0 6.2 8.1
D 8.0 7.1 8.5
Newborn Screening for Maple Syrup Urine Disease (MSUD)
Using a Totally Automated System.
Claudio Sampaio Filho (Jr)1, Fabiola Frias1, Raquel Veturiano1,
Vanessa Silva1, Bruna Gallo1, Fabio Bernardes1, Jonatas Boalento1,
Anakele Massi1, Josias Lira1,
1-INTERCIENTIFICA, S.J.Campos - Brasil
Nº PO
Contact:
Conclusions:
The enzymatic colorimetric method (NeoLISA®MSUD Kit) in a totally automated system (Nimbus
NeoLISA®) is an excellent alternative to meet the needs of the Brazilian Newborn Screening
Program or any Newborn Screening Program looking to implement MSUD testing without the high
cost involved with MS/MS. The use of MS/MS for MSUD and other diseases has been studied by
some labs in Brazil. High cost associated with the equipment and accessories, human resources,
infrastructure of the labs, and technical support are considered problematic and usually associated
with the negative perspective to introduce MSUD testing. The use of NeoLISA®MSUD in a totally
automated system (Nimbus NeoLISA®) meets the perspective proposed by the studies of Rede
DXB with development of newborn screening program that recognize the professional’s efforts to
prevent the deleterious consequences in non-treated MSUD patients.
Claudio Augusto Sampaio Filho
csampaio@intercientifica.com.br
Av. Sishima Hifumi, 2911 – Urbanova
Pq. Tec. UNIVAP – mod. 306-309
12244-000 S.J.Campos, SP
Brazil
www.intercientifica.com.br
ic@intercientifica.com.br
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