Sains Malaysiana 43(12)(2014):
1855–1863
Incidence of Cronobacter sakazakii in Powdered Infant
Formula Milk Available in Malaysia
(Kehadiran Cronobacter sakazakii dalam Susu Tepung Rumusan
Bayi yang Terdapat di Malaysia)
NORRAKIAH ABDULLAH SANI1*, MASOMEH GHASSEM1, ABDUL SALAM BABJI1,
UMA PRIYA KUPUSAMY2 & NORIZAN JAAFAR2
1School
of Chemical Sciences and Food Technology, Faculty of Science and Technology
Universiti Kebangsaan Malaysia, 43600
Bangi, Selangor, Malaysia
2Chemistry Department
of Malaysia, 46661 Petaling Jaya, Selangor, Malaysia
Diserahkan: 4 Ogos 2011/Diterima: 30 September 2014
ABSTRACT
Enterobacter sakazakii previously known as 'yellow-pigmented E.
cloacae' has been classified as a new genus 'Cronobacter'based
on taxonomic analysis and geno-and phenotypic evaluation. This pathogenic
organism has been associated with rare form of infant meningitis
and necrotizing enterocolitis (NEC) with high mortality rate (40-80%). Some
cases have been linked to the consumption of contaminated powdered
infant formula milk (PIF).
The objective of this study was to determine the presence of Cronobacter
spp. in PIF
sold in Malaysia. A selective chromogenic agar, Brilliance
Enterobacter sakazakii (DFI,
Oxoid), was used for detection of Cronobacter strains. Presumptive
Cronobacter isolates were identified using biochemical tests
(API 20E and MicrogenTM)
and molecular assays (SYBR Green Real-time PCR and 16S ribosomal DNA sequencing).
All presumptive Cronobacter strains produced typical blue-green
colonies and non-Cronobacter strains produced yellow colonies
on Brilliance Enterobacter sakazakii agar (DFI formulation). A total of 12 presumptive isolates
were selected from DFI agar and identified with biochemical and
molecular tests. The results indicated prevalence of 12.5% C.
sakazakii contamination from 72 PIF
samples. Molecular detection methods such as Real-time
PCR and
16S rDNA
proved to have higher identification percentage compared
to the biochemical tests. In this study, it was observed that molecular
assays were suitable means for sensitive identification of Cronobacter
strains in PIF samples.
Keywords: Biochemical test; chromogenic media; Cronobacter sakazakii; enterobacteriaceae;
Real time-PCR
ABSTRAK
Enterobacter sakazakii yang sebelum ini dikenali
sebagai 'E. cloacae berpigmen kuning' telah diklasifikasikan
sebagai genus baru 'Cronobacter' berdasarkan analisis takson
serta semakan secara fenotip dan genotip. Organisma patogen ini dikaitkan dengan penyakit meningitis pada
bayi dan enterokolitis nekrosis (NE) dengan kadar kematian
yang tinggi (40-80%). Sesetengah kes dikaitkan dengan pengambilan
susu tepung rumusan bayi (PIF) yang telah tercemar.
Objektif kajian ini adalah untuk menentukan kehadiran Cronobacter spp. di dalam PIF di Malaysia. Suatu agar
pilihan kromogenik, Brilliance Enterobacter sakazakii (DFI,
Oxoid) telah digunakan bagi mengenal pasti pencilan Cronobacter.
Pencilan Cronobacter jangkaan telah dikenal pasti dengan
menggunakan ujian biokimia (API 20E dan MicrogenTM) dan asai molekul (SYBR Hijau
Masa-nyata PCR
dan penjujukan 16S DNA ribosom). Semua pencilan Cronobacter
jangkaan telah menghasilkan koloni berwarna biru kehijauan manakala
pencilan bukan Cronobacter menghasilkan koloni berwarna kuning
di atas agar Brilliance Enterobacter sakazakii (DFI
formulation). Sejumlah 12 pencilan
jangkaan telah dipilih daripada agar DFI dan dikenal pasti dengan menggunakan ujian
biokimia dan molecular. Hasil kajian
menunjukkan kontaminasi lazim oleh C.
sakazakii daripada 72 sampel PIF
adalah sebanyak 12.5%. Kaedah
pengesanan molekular seperti Masa-nyata PCR dan 16S rDNA telah menunjukkan peratus
pengenalpastian yang lebih tinggi berbanding dengan hasil ujian
biokimia. Kajian ini menunjukkan pengasaian secara molekular
adalah suatu cara yang sesuai dan sensitif bagi mengenal pasti pencilan
Cronobacter dalam sampel PIF.
Kata kunci: Cronobacter
sakazakii; enterobacteriaceae; Masa-nyata PCR;
medium kromogenik; ujian biokimia
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*Pengarang
untuk surat-menyurat; email: norrasani@ukm.edu.my
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