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             \author[1]{Daissy Vargas  SepAlveda}

             \affil[1]{  University College of Cundinamarca}

\renewcommand\Authands{ and }

\date{\small \em Received: 13 December 2012 Accepted: 4 January 2013 Published: 15 January 2013}

\maketitle


\begin{abstract}
        


For many years the trichrome staining technique (TricrómicWheatley) has been considered as the most important technique for the identification ofthe most common intestinal protozoa and popularin parasitology (1).Currently the mostsensiblidad procedure for detecting and identifying protozoa trophozoites stool sample as it helps to easily highlight the morphology of amoebic cysts and trop-hozoites however, the procedure is complicated and tediousto perform and require at seven different reagents which is probably the most critical especially in laboratories with limited staff, this makes it complicated the routine use of this technique in most of the clinical laboratory, using koplic additionally facilitates reagent contamination by repeated use.(4,5

\end{abstract}


\keywords{tricrómicwheatley, diagnosis of amoebae.}

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\let\tabcellsep& 	 	 		 
\section[{Introduction}]{Introduction}\par
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.\par
Author: Bacteriologa, Universidad Colegio Mayor de Cundinamarca. Colombia, Afiliation: CMD SIPLAS. e-mail: daissyjasbeydi@gmail.com 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.(  {\ref 2}) 
\section[{Original Technical Steps}]{Original Technical Steps} 
\section[{Wheatley's Modification of the Gomori}]{Wheatley's Modification of the Gomori}\par
Note: formalin fixed Fecal samples are suitable for this dyeing process 
\section[{a) Important considerations}]{a) Important considerations}\par
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. (  {\ref 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.  
\section[{Sensitivity and Specificity}]{Sensitivity and Specificity}\par
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 
\section[{a) Parasitic forms identification with modified technique}]{a) Parasitic forms identification with modified technique}\par
Micrographs of amoebae obtained modified tech-nique implemented     \begin{figure}[htbp]
\noindent\textbf{1}\includegraphics[]{image-2.png}
\caption{\label{fig_0}Figure 1 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{2}\includegraphics[]{image-3.png}
\caption{\label{fig_1}Figure 2 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{3}\includegraphics[]{image-4.png}
\caption{\label{fig_2}Figure 3 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{4}\includegraphics[]{image-5.png}
\caption{\label{fig_3}Figure 4 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{5}\includegraphics[]{image-6.png}
\caption{\label{fig_4}Figure 5 :Conclusions?}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{1} \par 
\begin{longtable}{P{0.85\textwidth}}
Experimental development\\
Validation of the art Stian Modified Trichrome in Cmd\\
Siplas\end{longtable} \par
 
\caption{\label{tab_1}Table 1 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{DE} \par 
\begin{longtable}{P{0.06108829568788501\textwidth}P{0.46427104722792606\textwidth}P{0.04945242984257358\textwidth}P{0.09075975359342915\textwidth}P{0.016872005475701574\textwidth}P{0.025598904859685148\textwidth}P{0.04828884325804243\textwidth}P{0.07854209445585215\textwidth}P{0.015126625598904861\textwidth}}
\multicolumn{2}{l}{b) Kappa index leukocytes}\tabcellsep \multicolumn{3}{l}{TABLA DE 2*2}\tabcellsep \\
\tabcellsep \tabcellsep \tabcellsep \multicolumn{3}{l}{Reference}\tabcellsep No reference\\
\tabcellsep \tabcellsep \tabcellsep \multicolumn{3}{l}{Reagent}\tabcellsep Reagent\\
\tabcellsep \multicolumn{4}{l}{OBSERVER 1 Reagent to validate Vp}\tabcellsep \tabcellsep Fp\\
\tabcellsep \multicolumn{5}{l}{concordance No Reagent to validate FN}\tabcellsep VN\\
\tabcellsep \tabcellsep \tabcellsep \multicolumn{3}{l}{in identification Vp+FN}\tabcellsep negative VN+Fp\\
OBSERVADOR 2\tabcellsep \multicolumn{2}{l}{concordance in identifying parasitic forms Sensitivity Specificity negative}\tabcellsep \multicolumn{4}{l}{f l k 1 Vp/(Vp+FN)=True positives t 0 VN/(VN+Fp)=True Negatives CRITERIO DE 0 D 16 ACEPTABILIDA}\tabcellsep CRITERIO DE ACEPTABILIDAD\\
2\tabcellsep \multicolumn{3}{l}{17 \%Sensitivity ÍNDICE KAPPA VPP (\%)}\tabcellsep 1\tabcellsep \multicolumn{2}{l}{100,0}\tabcellsep 16 1,00 100,0\tabcellsep DIAGNOSTICACCURACYIS NOTED\\
\tabcellsep \multicolumn{2}{l}{\%Specificity VNP (\%)}\tabcellsep \tabcellsep \tabcellsep \multicolumn{2}{l}{100,0}\tabcellsep 100,0\\
\tabcellsep \multicolumn{3}{l}{Total Positives}\tabcellsep \tabcellsep \tabcellsep 1\\
Volume XIII Issue VII Version I\tabcellsep \multicolumn{6}{l}{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\tabcellsep \multicolumn{4}{l}{2*2 Reference Reagent ÍNDICE KAPPA Reagent to validate Vp VPP (\%)}\tabcellsep \multicolumn{2}{l}{No reference Reagent 1,00 Fp 100,0}\tabcellsep Aceptability DIAGNOSTICACCURACYIS NOTED\\
\tabcellsep \multicolumn{3}{l}{No Reagent to VNP (\%) validate Total positives FN}\tabcellsep \tabcellsep \tabcellsep 100,0 VN 1\\
\tabcellsep \multicolumn{4}{l}{Vp+FN Total Negatives}\tabcellsep \tabcellsep VN+Fp 16\\
\tabcellsep Sensitivity\tabcellsep \multicolumn{5}{l}{Vp/(Vp+FN)=True positives}\\
\tabcellsep Specificity\tabcellsep \multicolumn{5}{l}{VN/(VN+Fp)=True Negatives ÍNDICE KAPPA}\\
\tabcellsep \tabcellsep \tabcellsep \tabcellsep Pe\tabcellsep \tabcellsep 0,003\\
\tabcellsep \tabcellsep \tabcellsep \tabcellsep Po\tabcellsep \tabcellsep 1,00\tabcellsep Acceptability\\
c) Kappa index yeast\tabcellsep \tabcellsep \tabcellsep \tabcellsep \tabcellsep \\
\tabcellsep \multicolumn{3}{l}{ÍNDICE KAPPA}\tabcellsep \tabcellsep \tabcellsep 1,00\tabcellsep DIAGNOSTICACCURACYIS NOTED\\
\tabcellsep \tabcellsep \multicolumn{3}{l}{OBSERVER 1}\tabcellsep \\
\tabcellsep \multicolumn{2}{l}{VPP (\%)}\tabcellsep \multicolumn{4}{l}{100,0 concordance in}\\
\tabcellsep \multicolumn{2}{l}{VNP (\%)}\tabcellsep \tabcellsep \multicolumn{3}{l}{100,0 identification in}\tabcellsep negative\\
BSERVADOR 2\tabcellsep \multicolumn{6}{l}{Total positives Total Negatives Pe ÍNDICE KAPPA yeast 16 1 0,886 concordance in identifying parasitic forms 1 negative 0}\tabcellsep 0 16\\
\tabcellsep 17\tabcellsep \tabcellsep Po\tabcellsep \multicolumn{2}{l}{1}\tabcellsep 1,00\tabcellsep 16\\
\tabcellsep \%Sensitivity\tabcellsep \tabcellsep \tabcellsep \tabcellsep \tabcellsep 100,0\\
\tabcellsep \%Specificity\tabcellsep \tabcellsep \tabcellsep \tabcellsep \tabcellsep 100,0\end{longtable} \par
 
\caption{\label{tab_2}TABLE DE}\end{figure}
 			\footnote{© 2013 Global Journals Inc. (US) © 2013 Global Journals Inc. (US)} 			\footnote{© 2013 Global Journals Inc. (US)} 			\footnote{   ( )   } 		 		\backmatter  			  				\begin{bibitemlist}{1}
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\bibitem[Garcia and Shimizu (1998)]{b2}\label{b2} 	 		‘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 		.  	 	 		\textit{Journal of Clinical Microbiology}  		July 1998. 36  (7)  p. .  	 
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