Jurnal Reading Infection I

Friday, 21 December 2019

COMPARATIVE STUDY IN EARLY NEONATES WITH

SEPTICEMIA BY BLOOD CULTURE, STAINING
TECHNIQUES AND C - REACTIVE PROTEIN (CRP)

Ekarini Katharina Yunarti Nabu

Advisor :
Dr. dr Ni Nyoman Sri Budayanti, Sp.MK(K)
 INTRODUCTION
Def : clinical
syndrome occurring in
the first 30 days of life
due to infectious
agents

Remains a
Etiological agents SEPTICEMIA significant cause
of mortality in
vary in different
geographical area NEONATES developing
countries

Infection therefore
becomes fulminant
and causes death
within few hour or
days
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 INTRODUCTION

Acridine orange which is a fluorochrome stain,

demonstration of bacteria in blood and binding
with nucleic acid

CRP for detecting the early infection of

neonatal septicemia by using semiquantitative
latex –agglutination ( rapid screening)

This study to compare and evaluate the gram

stain, acridine orange stain and CRP result with
blood culture reports

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 MATERIAL AND METHODS

70 samples
Sick neonatal in government Rajaji Hospital Madurai India
December 2013 – September 2014
Inclusion Criteria Exclusion Criteria
Refusal of feeds Spina Bifida
Lethargi,Diarrhea Anencephaly
Respiratory distress Other neural tube defects and
Vomiting,poor cry ,convulsion in very sick neonates
Abdominal distention,fever
Hepatomegali jaundice,umbilical sepsis

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 C-reactive protein (CRP) test
(by TULIP Diagnostic)

Semiquantitative
Qualitative Method
Method

1 drop serum + 1 drop

Using isotonic saline
rhelax CRP latex reagent

Observing for agglutination

macroscopically after two Serial doubling dilutions of the
minutes serum sample was prepared as
follows 1:2,1:4,1:8, 1: 16,
1:32,1:64

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Buffy Coat Smear Examination : 1 ml blood sterile wintrobe
tubecentrifuge at 2500 rpm for 30 minutesplasma was
removed, buffycoat was taken-> smear was stained by gram’s
technique and acridine orange with fixed by methanol.

Buffy Coat Smear Examination

Gram’S Staining Acridine Orange Staining
1. Crystal Violet ( 1 menit) Left for 5 minutes
2. Iodine (1 menit) Examined with incident
3. Acetone ( 5 seconds) litght fluorescence 600x and
4. Carbol Fuchsin (1 menit) confirmed at 1500x magnification
Gram + violet appeared orange/orange red
Gram -  pink

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 Blood Culture Method
Blood Collected to a culture bottle containing brain heart
infusion broth
Incubated at 37 0C 18-24 hours, sub culture on plate

1. Blood Agar Plate 2. Mac Conkey Agar Plate (incubated 37

0C 18 -24 hours)

Colony size, shape,edge, Broth was examined for any

margin,consistency was noted turbidity, pellicle formation.

Identification by : Gram Stain, Hanging drop to find of motile &non motile

organism, Oxidase,catalase to identifity S.Aureus and coNS
Member of species was identified by biochemical reactions : sugar fermentation,
indol, methyl red, citrate, urease, hidrogern sulphide
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 Kirby Bauer- Disc Diffusion Technique

■ Organism antimicrobial sensitivity test was done to

select antimicrobial drugs
■ Identify the zone inhibition of bacterial strain
■ Used ampicillin,gentamycin, C0-
trimoxazole,cefotaxime,amikacin
■ Cloxacillin for gram positive agents

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RESULTS

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RESULTS

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RESULTS

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RESULTS

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 DISCUSSION

41 Of the 70 patients

37 (90.24 %) 4 (9.76 %)
Gram Negative Gram Positive
1. Klebsiella 28 (68.29 %) coNS
2. Pseudomonas Aeruginosa 3 (7.31%)
3. E.coli 4 (9.76 %)
4. Proteus Genus 2 (4.88 %)
This report is supported by study of Vaidya et al.

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 DISCUSSION

■ Gram Negative organisms like K. pneumonia,E coli, Proteus Mirabilis highly sensitive
to cefotaxime and amikacin.
■ Gentamycin was moderately sensitive to gram + and gram –
■ coNS was sensitive to amikacin, gentamycin,cloxacillin
(supported by Vaidya et al, Gupta et al)
■ In this study showing the sensitivity,specificity and PPV are 90.2 %, 100 % and 100%
( Sucilathangam et al)
■ In this study, that out of 70 cases septicemia, most the infant 62 (88.57%)belonged
to lower birth weight of less than 2.5 kg,41 % cases were blood culture positive and
29 % cases were blood culture negative (correlated with the study of Desai et al).

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 CONCLUSION

Male Neonates (54.29 %) were more

affected than female neonates (45.7%)

Serum CRP and Buffy Coat Smear

examination is simple method for
diagnosis of childhood septicemia.

The isolated organism were found to be

highly sensitive to cefotaxime and
amikacin

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