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  <front>
    <journal-meta>
      <journal-id journal-id-type="nlm-ta">Arq Bras Cardiol</journal-id>
      <journal-id journal-id-type="publisher-id">abc</journal-id>
      <journal-title-group>
        <journal-title>Arquivos Brasileiros de Cardiologia</journal-title>
        <abbrev-journal-title abbrev-type="publisher">Arq. Bras. Cardiol.</abbrev-journal-title>
      </journal-title-group>
      <issn pub-type="ppub">0066-782X</issn>
      <issn pub-type="epub">1678-4170</issn>
      <publisher>
        <publisher-name>Sociedade Brasileira de Cardiologia - SBC</publisher-name>
      </publisher>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v3">WH7L7sR94pn6WZcH7xDCZCL</article-id>
      <article-id pub-id-type="other">01092</article-id>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v2">S0066-782X2020001401092</article-id>
      <article-id pub-id-type="publisher-id">abc.20200656</article-id>
      <article-id pub-id-type="doi">10.36660/abc.20200656</article-id>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Minieditorial</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Testes Sorol&#243;gicos para Doen&#231;a de Chagas: Outra Evid&#234;ncia de Enigma em Doen&#231;a Amplamente Negligenciada</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0002-1090-8165</contrib-id>
          <name>
            <surname>Schmidt</surname>
            <given-names>Andr&#233;</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">
            <sup>1</sup>
          </xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Marin</surname>
            <given-names>Jos&#233; Antonio</given-names>
            <suffix>Neto</suffix>
          </name>
          <xref ref-type="aff" rid="aff1">
            <sup>1</sup>
          </xref>
        </contrib>
      </contrib-group>
      <aff id="aff1">
        <label>1</label>
        <institution content-type="orgname">Universidade de S&#227;o Paulo</institution>
        <institution content-type="orgdiv1">Faculdade de Medicina de Ribeir&#227;o Preto</institution>
        <institution content-type="orgdiv2">Centro de Cardiologia</institution>
        <addr-line>
          <named-content content-type="city">Ribeir&#227;o Preto</named-content>
          <named-content content-type="state">SP</named-content>
        </addr-line>
        <country country="BR">Brasil</country>
        <institution content-type="original">Universidade de S&#227;o Paulo Faculdade de Medicina de Ribeir&#227;o Preto - Centro de Cardiologia, Ribeir&#227;o Preto, SP &#8211; Brasil</institution>
      </aff>
      <author-notes>
        <corresp id="c01"> Correspond&#234;ncia: Andr&#233; Schmidt &#8226; Universidade de S&#227;o Paulo Faculdade de Medicina de Ribeir&#227;o Preto - Centro de Cardiologia - HCFMRP-USP - Av. Bandeirantes, 3900. CEP 14048-900, Ribeir&#227;o Preto, SP &#8211; Brasil E-mail: <email>aschmidt@fmrp.usp.br</email>
				</corresp>
      </author-notes>
      <pub-date date-type="pub" publication-format="electronic">
        <day>18</day>
        <month>01</month>
        <year>2021</year>
      </pub-date>
      <pub-date date-type="collection" publication-format="electronic">
        <month>12</month>
        <year>2020</year>
      </pub-date>
      <volume>115</volume>
      <issue>6</issue>
      <fpage>1092</fpage>
      <lpage>1093</lpage>
      <permissions>
        <license license-type="open-access" xlink:href="https://creativecommons.org/licenses/by/4.0/" xml:lang="en">
          <license-p> This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. </license-p>
        </license>
      </permissions>
      <kwd-group xml:lang="pt">
        <kwd>Doen&#231;a de Chagas</kwd>
        <kwd>Epidemiologia</kwd>
        <kwd>Testes Sorol&#243;gicos</kwd>
        <kwd>Interven&#231;&#227;o Coron&#225;ria Percut&#226;nea/m&#233;todos</kwd>
      </kwd-group>
      <counts>
        <fig-count count="0"/>
        <table-count count="0"/>
        <equation-count count="0"/>
        <ref-count count="16"/>
        <page-count count="2"/>
      </counts>
    </article-meta>
  </front>
  <body>
    <p>Cardiologistas e Gastroenterologistas atuando em ambientes variados da Faculdade de Medicina da USP em Ribeir&#227;o Preto frequentemente deparam-se com pacientes apresentando caracter&#237;sticas epidemiol&#243;gicas, cl&#237;nicas e laboratoriais sugestivas da doen&#231;a de Chagas (DC), mas os testes sorol&#243;gicos espec&#237;ficos s&#227;o negativos.<sup><xref ref-type="bibr" rid="B1">1</xref></sup> Em s&#233;rie de 65 pacientes estudados de 01/07/2011 a 31/12/2012 com coronariografia invasiva para esclarecer o sintoma cardinal de dor precordial, todos com altera&#231;&#245;es de mobilidade segmentar ventricular (incluindo 28 com o t&#237;pico aneurisma apical na ventriculografia de contraste radiol&#243;gico) a sorologia com dois testes foi positiva em apenas 11(17%) dos casos.<sup><xref ref-type="bibr" rid="B2">2</xref></sup> Como elucidar esse enigma, e como conduzir tais casos? A resposta talvez esteja, em parte, na experi&#234;ncia com esta entidade, e no conhecimento e convic&#231;&#227;o sobre a acuidade diagn&#243;stica dos testes sorol&#243;gicos empregados.</p>
    <p>Mas o problema da quest&#227;o se inicia com a assimetria conceitual que paira sobre a incerteza envolvendo o diagn&#243;stico rotineiro de DC pela verifica&#231;&#227;o sorol&#243;gica de anticorpos antiparasit&#225;rios. Assim, de acordo com diretrizes aceitas,<sup><xref ref-type="bibr" rid="B3">3</xref> , <xref ref-type="bibr" rid="B4">4</xref></sup> enquanto um teste negativo seria suficiente para afastar o diagn&#243;stico (inclusive autorizando doa&#231;&#227;o de sangue ou &#243;rg&#227;os s&#243;lidos), a necessidade de dois testes sorol&#243;gicos positivos com m&#233;todos distintos para confirma&#231;&#227;o diagn&#243;stica da DC foi h&#225; muito reconhecida e implementada em rotinas cl&#237;nicas e de pesquisa; tal se justificaria devido &#224; heterog&#234;nea acur&#225;cia (m&#233;dia da sensibilidade e especificidade) dos testes, ex. fixa&#231;&#227;o de complemento, hemaglutina&#231;&#227;o indireta, imunofluoresc&#234;ncia indireta e aglutina&#231;&#227;o direta).<sup><xref ref-type="bibr" rid="B5">5</xref></sup> Mesmo com o surgimento da quimioluminesc&#234;ncia baseada em ELISA, m&#233;todo automatizado para eliminar o componente subjetivo daqueles testes acima citados, a recomenda&#231;&#227;o de utilizar dois testes distintos simultaneamente, seguido de um terceiro em caso de discord&#226;ncia inicial, persiste na recomenda&#231;&#227;o da Organiza&#231;&#227;o Panamericana de Sa&#250;de.<sup><xref ref-type="bibr" rid="B6">6</xref></sup> Contraditoriamente, nota t&#233;cnica da Organiza&#231;&#227;o Mundial da Sa&#250;de recomendou a utiliza&#231;&#227;o de um &#250;nico teste baseado em ELISA, por quimioluminesc&#234;ncia para &#8220;screening&#8221; em bancos de sangue, apoiada na alegada muito alta sensibilidade (seria pr&#243;ximo de 99%), e com o intuito de reduzir custos ao realizar grande volume de testes de triagem em hemocentros e demais bancos de sangue.<sup><xref ref-type="bibr" rid="B7">7</xref></sup> Contudo, h&#225; evid&#234;ncias bem documentadas de que tal protocolo possa ser inadequado.<sup><xref ref-type="bibr" rid="B8">8</xref></sup></p>
    <p>O estudo publicado<sup><xref ref-type="bibr" rid="B9">9</xref></sup> , apresenta dados oriundos de hemocentros no Estado do Cear&#225; relativos a sorologia para DC ao longo de extenso per&#237;odo temporal e reaviva a evid&#234;ncia suscitada por algumas reflex&#245;es inquietantes:<sup><xref ref-type="bibr" rid="B10">10</xref></sup></p>
    <list list-type="order">
      <list-item>
        <p>Considerando ser essa regi&#227;o end&#234;mica, com transmiss&#227;o vetorial ainda ativa pelo pr&#243;prio relato dos Autores, chama aten&#231;&#227;o o elevado n&#250;mero de exames inconclusivos (70%) nos considerados doadores ineleg&#237;veis, e levantando a possibilidade de rea&#231;&#227;o cruzada com <italic>leishmania spp</italic> , tamb&#233;m end&#234;mica na regi&#227;o, e que poderia ser respons&#225;vel, pelo menos em parte, por resultados t&#227;o intrigantes.<sup><xref ref-type="bibr" rid="B11">11</xref></sup> Mas &#233; importante, desde j&#225;, enfatizar que pelo relato dos Autores, a inconclusividade inicial foi decorrente de um &#250;nico teste ELISA que n&#227;o seria nem positivo nem negativo. Isso &#233; diferente de inconclusividade decorrente de dois testes sendo um positivo e outro negativo, situa&#231;&#227;o reconhecida habitualmente em diretrizes.</p>
      </list-item>
      <list-item>
        <p>Caracteristicamente no trabalho em pauta, a repeti&#231;&#227;o do mesmo teste baseado em ELISA em segunda coleta mostrou que de 1.225 testes positivos ou inconclusivos pelo primeiro teste, em apenas 425 pacientes os resultados persistiam positivos ou inconclusivos. Isso sugere baixa reprodutibilidade, algo preocupante, e indiretamente corroborado pelo relato de que 43% dos exclu&#237;dos no primeiro exame j&#225; haviam sido doadores. Assim, a negatividade no exame de triagem da doa&#231;&#227;o anterior teria levado a potencial recep&#231;&#227;o de material oriundo de paciente &#8220;falso-negativo&#8221; para doen&#231;a de Chagas.</p>
      </list-item>
      <list-item>
        <p>Ainda, destes 425 pacientes dentre os 1.225 com resultados positivos (333) ou inconclusivos (92) persistindo ap&#243;s os dois primeiros testes baseados em ELISA na primeira e segunda amostras, foram escrutinizados outra vez nesta segunda amostra sangu&#237;nea com nova sorologia, desta feita com diferente m&#233;todo sorol&#243;gico (imunofluoresc&#234;ncia ou <italic>Western blot</italic> ), verificou-se que 305 foram negativos, 48 inconclusivos, e apenas 72 persistiram com testes positivos com base no terceiro exame sorol&#243;gico.</p>
      </list-item>
      <list-item>
        <p>Ent&#227;o, apenas houve concord&#226;ncia em 28% dos casos, cotejando-se os primeiros e os &#250;ltimos testes. Afinal, os doadores assim aprovados s&#227;o ou n&#227;o portadores de DC? E os rejeitados como doadores, que probabilidade b&#225;sica de DC possuem?</p>
      </list-item>
    </list>
    <p>Verifica-se, portanto, n&#237;tida necessidade de aprimoramento no diagn&#243;stico sorol&#243;gico, pois a transmiss&#227;o transfusional em nosso meio tem incid&#234;ncia incerta, por conta da paucidade de sintomas da fase aguda da DC, que muitas vezes se manifesta como quadro febril inespec&#237;fico, n&#227;o incomum em pacientes hospitalizados. A transi&#231;&#227;o para a forma indeterminada e a evolu&#231;&#227;o ulterior para formas cl&#237;nicas &#233; tardia e pode n&#227;o ser identificada se n&#227;o forem feitas buscas ativas. Em tempos remotos descreveu-se a ocorr&#234;ncia de contamina&#231;&#227;o em 6 (25%) de 24 casos que receberam transfus&#227;o contaminada, tendo um caso desenvolvido sinais cl&#237;nicos de miocardite na fase aguda.<sup><xref ref-type="bibr" rid="B12">12</xref></sup> Mais recentemente, casos graves foram descritos em outros pa&#237;ses.<sup><xref ref-type="bibr" rid="B13">13</xref></sup> No que tange &#224; transmiss&#227;o associada a transplantes de &#243;rg&#227;os s&#243;lidos infectados, com a concomitante imunossupress&#227;o, n&#227;o s&#227;o infrequentes manifesta&#231;&#245;es graves e potencialmente letais na fase aguda.<sup><xref ref-type="bibr" rid="B14">14</xref></sup> Al&#233;m disso, tamb&#233;m pode se dar reativa&#231;&#227;o da doen&#231;a em receptor insuspeito de portar DC, quando a aus&#234;ncia de um teste padr&#227;o-ouro para diagn&#243;stico contribui para as ocorr&#234;ncias mencionadas.</p>
    <p>A caracteriza&#231;&#227;o pela Organiza&#231;&#227;o Mundial da Sa&#250;de da DC como &#8220;negligenciada&#8221; adquire certamente mais for&#231;a com a constata&#231;&#227;o de que, 50 anos ap&#243;s o estabelecimento de testes obrigat&#243;rios para doadores em nosso meio, ainda se verifica o descarte de bolsas por conta do elevado n&#250;mero de testes inconclusivos. Os impactos sociais e financeiros desta &#8220;inconsist&#234;ncia&#8221; sorol&#243;gica est&#227;o para serem determinados.</p>
    <p>De toda forma, essas duas vertentes do dilema imposto pela incerteza s&#227;o imanentes ao contexto de situa&#231;&#245;es enigm&#225;ticas envolvendo diagn&#243;stico quando inexiste padr&#227;o-ouro, e a acur&#225;cia de todo teste empregado no &#8220;mundo-real&#8221; v&#225; ser aferida em ambiente amostral diverso daquele em que o teste foi padronizado. Assim, persiste por se comprovar cabalmente a corre&#231;&#227;o da sufici&#234;ncia de apenas um teste negativo de alta sensibilidade para excluir a DC.<sup><xref ref-type="bibr" rid="B15">15</xref></sup> Al&#233;m disso, mesmo o algoritmo sucessivo (teste positivo de alta sensibilidade seguido de outro, com alta especificidade) para comprova&#231;&#227;o diagn&#243;stica<sup><xref ref-type="bibr" rid="B16">16</xref></sup> parece desafiado por evid&#234;ncias como as agora publicadas nos ABC por pesquisadores do Cear&#225;.</p>
  </body>
  <back>
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    <fn-group>
      <fn fn-type="other">
        <p>Minieditorial referente ao artigo: Preval&#234;ncia da Infec&#231;&#227;o pelo Trypanosoma cruzi em Doadores de Sangue</p>
      </fn>
    </fn-group>
  </back>
  <sub-article article-type="translation" id="TRen" xml:lang="en">
    <front-stub>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Short Editorial</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Serological Tests in Chagas Disease: Another Enigmatic Evidence in a Disease Largely Neglected</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <contrib-id contrib-id-type="orcid">0000-0002-1090-8165</contrib-id>
          <name>
            <surname>Schmidt</surname>
            <given-names>Andr&#233;</given-names>
          </name>
          <xref ref-type="aff" rid="aff1001">
            <sup>1</sup>
          </xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Marin</surname>
            <given-names>Jos&#233; Antonio</given-names>
            <suffix>Neto</suffix>
          </name>
          <xref ref-type="aff" rid="aff1001">
            <sup>1</sup>
          </xref>
        </contrib>
      </contrib-group>
      <aff id="aff1001">
        <label>1</label>
        <country country="BR">Brazil</country>
        <institution content-type="original">Universidade de S&#227;o Paulo Faculdade de Medicina de Ribeir&#227;o Preto - Centro de Cardiologia, Ribeir&#227;o Preto, SP &#8211; Brazil</institution>
      </aff>
      <author-notes>
        <corresp id="c01001"><bold>Mailing Address: Andr&#233; Schmidt</bold> &#8226; Universidade de S&#227;o Paulo Faculdade de Medicina de Ribeir&#227;o Preto - Centro de Cardiologia - HCFMRP-USP - Av. Bandeirantes, 3900. Postal Code 14048-900, Ribeir&#227;o Preto, SP &#8211; Brazil E-mail: aschmidt@fmrp.usp.br</corresp>
      </author-notes>
      <kwd-group xml:lang="en">
        <kwd>Chagas Disease</kwd>
        <kwd>Epidemiology</kwd>
        <kwd>Sorologic Tests</kwd>
        <kwd>Percutaneous Coronary Intervention/methods</kwd>
      </kwd-group>
    </front-stub>
    <body>
      <p>Cardiologists and Gastroenterologists acting in distinct environments of our institution in Ribeir&#227;o Preto, not infrequently face patients with epidemiologic, clinical and laboratorial features of Chagas Disease (CD) in whom serological evaluation is negative for the disease.<sup><xref ref-type="bibr" rid="B1">1</xref></sup> In a series of 65 patients evaluated between 07/01/2011 and 12/31/2012 by invasive coronary angiography to elucidate the cardinal symptom of chest pain, all with wall motion abnormalities (including 28 presenting the typical apical aneurysm at contrast ventriculography), two distinct serological tests were positive in only 11(17%) patients.<sup><xref ref-type="bibr" rid="B2">2</xref></sup> How can we elucidate this enigma and provide guidance in this situation? The answer may be, in part, in the experience with this entity and in the knowledge and confidence in the diagnostic accuracy of the serological tests employed.</p>
      <p>The hitch point begins with the asymmetrical concepts leading to uncertainty that involve the routine diagnostic procedures of CD by using serological verification of antiparasite antibodies. In accordance to current guidelines,<sup><xref ref-type="bibr" rid="B3">3</xref> , <xref ref-type="bibr" rid="B4">4</xref></sup> only one negative test is enough to exclude the diagnosis (granting blood donation or solid organ transplantation), while two distinct positive serological tests are needed to confirm CD diagnosis. This is an ancient practice in clinical and research routines, due to the heterogeneous accuracy (mean of sensibility and specificity) of the tests/ e. g. complement fixation, indirect hemagglutination, indirect immunofluorescence and direct agglutination).<sup><xref ref-type="bibr" rid="B5">5</xref></sup> Even after the development of chemiluminescence methods based on ELISA, capable of automated reading to avoid subjective interpretation as seen in the above-mentioned tests, the rule of two simultaneous distinct tests followed by a third one if a discordant result is obtained, remains in the latest PAHO guidelines for diagnosis of CD.<sup><xref ref-type="bibr" rid="B6">6</xref></sup> A conflicting technical note from the WHO recommended a single ELISA test for screening in blood banks, supported by its alleged high sensitivity (nearly 99%) and as a way of cost reduction when dealing with a high volume of screening as occurs in blood centers and blood banks.<sup><xref ref-type="bibr" rid="B7">7</xref></sup> Nevertheless, well-documented evidence exists suggesting that this protocol may not be entirely adequate.<sup><xref ref-type="bibr" rid="B8">8</xref></sup></p>
      <p>The study published in this issue of Arquivos Brasileiros de Cardiologia,<sup><xref ref-type="bibr" rid="B9">9</xref></sup> presents data from blood centers in Cear&#225; state related to CD serological tests in an extensive period and renew the evidence leading to some unsettling reflections:<sup><xref ref-type="bibr" rid="B10">10</xref></sup></p>
      <list list-type="order">
        <list-item>
          <p>Since it is an endemic region with vectorial transmission still present by the Authors report, the strikingly high number of inconclusive tests (70%) in those donors considered ineligible, rises the possibility of a cross reaction with <italic>leishmania spp</italic> , also endemic in the region, that could be responsible, at least partially, for those so intriguing results.<sup><xref ref-type="bibr" rid="B11">11</xref></sup> It is relevant, from now on, to emphasize that from the Authors report, the initial inconclusive result was based in only one ELISA neither positive nor negative. This is distinct from an inconclusive result of two tests, one positive and other negative, a common finding in clinical guidelines.</p>
        </list-item>
        <list-item>
          <p>Characteristically in the present paper, the repetition of the ELISA test in a second sample in 1,225 previously positive or inconclusive results, only in 425 samples remained positive or inconclusive. This result suggests low reproducibility, a concerning result reinforced by their report that 43% of those excluded in the first test had previously donated blood. This indicates that the negative result in the first screening test led to the potential acceptance of a CD false-negative donor.</p>
        </list-item>
        <list-item>
          <p>Furthermore, those 425 donors with positive (333) or inconclusive (92) results after two ELISA tests in two distinct samples, had their second sample submitted to another serological test (immunofluorescence or <italic>Western blot</italic> ). In this third test, 305 were negative, 48 inconclusive and only 72 remained positive.</p>
        </list-item>
        <list-item>
          <p>In summary, there was a concordance of only 28% between the first and last tests. Can we state that those approved as donors are free of CD? What about those excluded as donors, what is their basic probability of having the disease?</p>
        </list-item>
      </list>
      <p>There is clearly the necessity of improvement in serological diagnostic tests since transfusional transmission of CD in our country remains uncertain, due to the paucity of symptoms in the acute phase of CD, mainly a non-specific febrile condition commonly seen in hospitalized patients. The transition to the indeterminate form and its ulterior clinical forms occurs lately and may not be identified if not actively scrutinized. In ancient times a description of transfusional contamination in 6 (25%) of 24 recipients of one contaminated bag had only one case with clinical signs of the acute phase myocarditis.<sup><xref ref-type="bibr" rid="B12">12</xref></sup> Recently, severe cases have been described in other countries.<sup><xref ref-type="bibr" rid="B13">13</xref></sup> When occurring in solid organ transplants recipients, due to the concomitant immunosuppression, it is not rare severe and potentially lethal manifestations of the acute phase of CD.<sup><xref ref-type="bibr" rid="B14">14</xref></sup> In addition, reactivation of the disease may occur in an organ recipient not suspected of having CD. The absence of a gold-standard diagnostic test contributes to the above occurrences.</p>
      <p>The definition by the World Health Organization of Chagas disease as &#8220;neglected&#8221; certainly acquires more strength by the recognition that 50 years after the establishment of mandatory tests in blood donors in our country, the number of discarded bags due to the number of inconclusive results is huge. Financial and social impact of this serological inconsistency are still to be established.</p>
      <p>Anyway, these two pathways of the dilemma imposed by uncertainty are immanent to the context of enigmatic situations involving diagnosis when no gold-standard exists and accuracy of every test applied to the &#8220;real-world&#8221; will be assessed in a diverse environment from the one it was standardized. Therefore, it rests to be properly established the sufficiency of only one high sensibility serological test to exclude CD.<sup><xref ref-type="bibr" rid="B15">15</xref></sup> Besides, even a two-step algorithm (a positive high sensibility test followed by a high specificity one) to establish diagnosis<sup><xref ref-type="bibr" rid="B16">16</xref></sup> seems challenged by evidence now published by researches from the state of Cear&#225;.</p>
    </body>
    <back>
      <fn-group>
        <fn fn-type="other">
          <p>Minieditorial referente ao artigo: Preval&#234;ncia da Infec&#231;&#227;o pelo Trypanosoma cruzi em Doadores de Sangue</p>
        </fn>
      </fn-group>
    </back>
  </sub-article>
</article>
