1. Introduction

he main purpose of this study was to evaluatea new method for obtaining atrichromic staining faster and effective for it used the samedyesand proce-ededim plementing two different technical koplicone with and one with direct drops of reagent in the lamina.

Author: Bacteriologa, Universidad Colegio Mayor de Cundinamarca. Colombia, Afiliation: CMD SIPLAS. e-mail: [email protected] There were 20 positive smearsall parasites and made several technical modifications in order to simplify and expedite the procedure equally maintaining the excellent staining qualities, the nimplemented the steps mentioned in the original technique and then the technique modified. ethanol should be as free of water as possible to avoid both the reactive evaporation of moisture absorption as that can preventeasy identification of the parasite.( 2)

2. Original Technical Steps

3. Wheatley's Modification of the Gomori

Note: formalin fixed Fecal samples are suitable for this dyeing process

4. a) Important considerations

The fund continues to see green and cytoplasm of protozoa is stained a blue green and purple. There dnuclei with inclusions purple and intracellular structures are easy to distinguish as glycogen vacuoles are the Iodamoeba butschlii. ( 6 Kappa: the agreement between observers for the identification of parasitic forms, leukocytes, yeasts, and negative for themis 1.0, which shows diagnostic accuracy and level of agreement between observers for the samples with the latest changes made by SIPLAS medical laboratory, concluding that the changes mentioned here allow adequate identification of both parasite forms leukocytes, yeast and other fungal forms structures that allow the definition diagnosed patients, ensuring diagnostic accuracy versus the clinical definition kappa Degree of agreement < 0 without agreement 0 -0.

5. Sensitivity and Specificity

The sensitivity and specificity of the samples analyzed for fungal structures, yeast and parasiticleucoidesis 100%, which shows that the stain can classify patients according to the irpositive or negative real state against it sclinical definition

6. a) Parasitic forms identification with modified technique

Micrographs of amoebae obtained modified tech-nique implemented

Figure 1 : Cysts Blastocystis hominis — Figure 1. Figure 1 :

Figure 2 : Cysts Entamoeba coli — Figure 2. Figure 2 :

Figure 3 : Cysts Iodamoeba butschlii — Figure 3. Figure 3 :

Figure 4 : Cysts Endolimax nana — Figure 4. Figure 4 :

Figure 5 : Cysts Giardia Duodenalis Note: The photomicrographs were taken by the bacteriologys Daissy Vargas Sepulveda in CMD SIPLAS IV. — Figure 5. Figure 5 :Conclusions?

Figure 6. Table 1 :

Experimental development

Validation of the art Stian Modified Trichrome in Cmd

Siplas

Figure 7. TABLE DE

b) Kappa index leukocytes		TABLA DE 2*2
			Reference			No reference
			Reagent			Reagent
	OBSERVER 1 Reagent to validate Vp					Fp
	concordance No Reagent to validate FN					VN
			in identification Vp+FN			negative VN+Fp
OBSERVADOR 2	concordance in identifying parasitic forms Sensitivity Specificity negative		f l k 1 Vp/(Vp+FN)=True positives t 0 VN/(VN+Fp)=True Negatives CRITERIO DE 0 D 16 ACEPTABILIDA				CRITERIO DE ACEPTABILIDAD
2	17 %Sensitivity ÍNDICE KAPPA VPP (%)			1	100,0		16 1,00 100,0	DIAGNOSTICACCURACYIS NOTED
	%Specificity VNP (%)				100,0		100,0
	Total Positives					1
Volume XIII Issue VII Version I	TABLA DE 2*2 Reference Reagent Reagent to validate Vp Vp+FN Sensitivity Vp/(Vp+FN)=True positives No reference Reagent Fp VN+Fp Specificity VN/(VN+Fp)=True Negatives Total Negatives 16 ÍNDICE KAPPA Pe 0,003 No Reagent to validate FN VN Po 1,00
( ) K	2*2 Reference Reagent ÍNDICE KAPPA Reagent to validate Vp VPP (%)				No reference Reagent 1,00 Fp 100,0		Aceptability DIAGNOSTICACCURACYIS NOTED
	No Reagent to VNP (%) validate Total positives FN					100,0 VN 1
	Vp+FN Total Negatives					VN+Fp 16
	Sensitivity	Vp/(Vp+FN)=True positives
	Specificity	VN/(VN+Fp)=True Negatives ÍNDICE KAPPA
				Pe		0,003
				Po		1,00	Acceptability
c) Kappa index yeast
	ÍNDICE KAPPA					1,00	DIAGNOSTICACCURACYIS NOTED
		OBSERVER 1
	VPP (%)		100,0 concordance in
	VNP (%)			100,0 identification in			negative
BSERVADOR 2	Total positives Total Negatives Pe ÍNDICE KAPPA yeast 16 1 0,886 concordance in identifying parasitic forms 1 negative 0						0 16
	17		Po	1		1,00	16
	%Sensitivity					100,0
	%Specificity					100,0

Appendix A

Diagnóstico Parasitología Médica, 5 ªed, 2009. Washington, DC 4. García, LS: ASM Press.
Diagnostic Medical Parasitology, L S Garcia , D A Bruckner . 1993. Washington D.C: American Society for Microbiology. (2nd ed)
Evaluation of Intestinal Protozoan Morphology in Human Fecal Specimens Preserved in Eco Fix: Comparison of Wheatley's Trichrome Stain and Eco Stain. Lynne S Garcia , Robyn Y Shimizu . Journal of Clinical Microbiology July 1998. 36 (7) p. .
Selección y Uso de Procedimientos de laboratorio para El diagnóstico de infecciones parasitarias deel tracto gastrointestinal. Normas De Laboratorio Clínico Instituto , P A Villanova , García , Ls . Cumitech 2003. 2007. ASM Press. 30.
Procedimientos para la recuperación e identificación de parásitos en el tracto intestinal, Directrices aprobadas, (edimientos para la recuperación e identificación de parásitos en el tracto intestinal, Directrices aprobadas) 2005. p. . Normas de Laboratorio Clínico Institute
PrácticaGuía para el Diagnóstico de Parasitología, 2 ª ed, Washington, DC: ASM Press.
Un procedimiento de tinción rápida para intestinal amebas y flagelados. Wheatley , De . Am. J. Clin. Pathol 1951. 21 p. .

Table of contents

Appendix A