Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Influenza is a contagious respiratory illness caused by influenza viruses A and B in humans and causes a significant amount of morbidity and mortality ...

Memorial Sloan Kettering Cancer Center

Cytomegalovirus (CMV) is a major threat in patients undergoing allogeneic bone marrow transplantation. The adoptive transfer of CMV-specific cytotoxic T lymphocytes (CTLs) expanded from the blood of CMV-seropositive donors has been shown to effectively control CMV infection. However, the requirement for safe and effective antigen-presenting cells (APCs) for each patient precludes broad applicability of this successful form of therapy. Here we analyze the ability of artificial APCs (AAPCs) to activate and expand CMV-specific CTLs from peripheral blood of seropositive HLA A2.1+ donors. We demonstrate that AAPCs expressing the CMV P495 peptide or the full-length pp65 protein stimulate P495-specific CTLs at least as effectively as autologous, peptide-pulsed, peripheral blood mononuclear cells or EBV-transformed B cells. Starting from 100 mL of blood, the AAPCs reliably yield clinically relevant CTL numbers after a single stimulation. CTLs activated on AAPCs effectively kill CMV-infected fibroblasts and have a Tc1 memory effector phenotype identical to that of CTLs generated with autologous APCs. AAPCs thus offer a rapid, controlled, convenient, and highly reproducible system for expanding CMV-specific CTLs. Furthermore, the CTL expansion obtained with AAPCs encoding full-length pp65 indicates that AAPCs may be used to present known as well as unknown CTL epitopes in the context of the AAPC's HLA.

Rapid expansion of cytomegalovirus-specific cytotoxic T lymphocytes by artificial antigen-presenting cells expressing a single HLA allele.

Human metapneumovirus is a recently discovered RNA virus that typically causes respiratory disease in children. It has been linked to severe lower airway disease in hematopoietic stem cell and solid-organ transplant recipients. hMPV infection in a large population of patients with underlying cancer and varying degrees of immunosuppression has not been reported. We sought to characterize hMPV infection in patients with cancer.

Clinical characterization of human metapneumovirus infection among patients with cancer.

Although usually mild, 2009 H1N1 Influenza has caused up to 6000 deaths in the US. To determine outcome in patients with cancer and/or hematopoietic stem cell transplant (HSCT), we reviewed our recent experience at Memorial Sloan-Kettering Cancer Center (MSKCC).

2009 H1N1 influenza infection in cancer patients and hematopoietic stem cell transplant recipients.

Effect of varicella-zoster virus (VZV) fluorescent-antibody-to-membrane-antigen (FAMA) testing on sensitivity of determining VZV immunity in healthcare workers and on furlough days.

&#x96BE;&#x8FA8;&#x68AD;&#x83CC;&#x662F;&#x8179;&#x6CFB;&#x7684;&#x536B;&#x751F;&#x4FDD;&#x5065;&#x76F8;&#x5173;&#x6700;&#x5E38;&#x89C1;&#x7684;&#x539F;&#x56E0;&#x3002;&#x51C6;&#x786E;&#x3001; &#x5FEB;&#x901F;&#x7684;&#x8BCA;&#x65AD;&#xFF0C;&#x6539;&#x5584;&#x75C5;&#x4EBA;&#x7684;&#x540E;&#x679C;&#x53CA;&#x9632;&#x6B62;&#x75BE;&#x75C5;&#x4F20;&#x64AD;&#x81F3;&#x5173;&#x91CD;&#x8981;&#x3002;&#x6211;&#x4EEC;&#x6BD4;&#x8F83;&#x6211;&#x4EEC;&#x4E24;&#x6B65;&#x8BCA;&#x65AD;&#x7B97;&#x6CD5;&#x540E;, &#x8DDF;&#x7684;&#x5468;&#x8F6C;&#x65F6;&#x95F4;&#x4E3A; 24 &#x5230; 48 &#x5C0F;&#x65F6;&#xFF0C;&#x4E0E; Cepheid Xpert C.&#x96BE;&#x8FA8;&#x8BA1;&#x5212;&#x514D;&#x75AB;&#x68C0;&#x6D4B;&#x4E2D;&#xFF0C;&#x7EC6;&#x80DE;&#x6BD2;&#x7D20;&#x4E2D;&#x548C;&#x8BD5;&#x9A8C; (&#x5E02;&#x957F;&#x8FD0;&#x901A;) pcr &#x7684;&#x5468;&#x8F6C;&#x65F6;&#x95F4;&#x4E0E;&#x8C37;&#x6C28;&#x9178;&#x8131;&#x6C22;&#x9176; (&#x7CA4;&#x6D77;) &#x9176;&#x8054;&#x514D;&#x75AB; &lt; 1 h&#x3002;&#x5728;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#x7684;&#x7B2C;&#x4E00;&#x9636;&#x6BB5;&#xFF0C;&#x53EA;&#x6709;&#x7CA4;&#x6D77;&#x9633;&#x6027;&#x7CAA;&#x4FBF;&#x6837;&#x672C;&#x7ECF;&#x8FC7;&#x5E02;&#x957F;&#x8FD0;&#x901A;&#x548C; Xpert PCR&#x3002;&#x7531;&#x4EA7;&#x6BD2;&#x6587;&#x5316;&#x89E3;&#x51B3;&#x4E0D;&#x4E00;&#x81F4;&#x7684;&#x7ED3;&#x679C;&#x3002;&#x5728;&#x7B2C;&#x4E8C;&#x9636;&#x6BB5;&#xFF0C;&#x6240;&#x6709;&#x7684;&#x7CAA;&#x4FBF;&#x6837;&#x672C;&#x7ECF;&#x8FC7;&#x7CA4;&#x6D77;&#x548C; Xpert PCR&#x3002;&#x53EA;&#x6709;&#x7CA4;&#x6D77;&#x9633;&#x6027;&#x5927;&#x4FBF;&#x7531;&#x5E02;&#x957F;&#x8FD0;&#x901A;&#x8FDB;&#x884C;&#x4E86;&#x8FDB;&#x4E00;&#x6B65;&#x7684;&#x6D4B;&#x8BD5;&#x3002;45 Xpert PCR &#x9633;&#x6027;&#x5927;&#x4FBF;&#x7684;&#x57FA;&#x56E0;&#x578B;&#x8868;&#x5F81;&#x8FDB;&#x884C; PCR &#x6838;&#x7CD6;&#x4E0E;&#x6280;&#x5408;&#x57FA;&#x56E0;&#x7684;&#x6D4B;&#x5E8F;&#x3002;&#x5728;&#x7B2C;&#x4E00;&#x9636;&#x6BB5;&#xFF0C;&#x7CA4;&#x6D77; &#xFF08;cyt&#xFF09; &#x4E0E;&#x7CA4;&#x6D77; Xpert PCR &#x7684;&#x534F;&#x8BAE;&#x4E3A; 72%&#x3002;&#x654F;&#x611F;&#x6027;&#x548C;&#x7CA4;&#x6D77;&#x5E02;&#x957F;&#x8FD0;&#x901A;&#x548C;&#x7CA4;&#x6D77; Xpert &#x805A;&#x5408;&#x9176;&#x94FE;&#x53CD;&#x5E94;&#x7684;&#x7279;&#x70B9;&#x5206;&#x522B;&#x662F; 57%&#x548C; 97%&#x548C; 100%&#x548C; 97%&#x3002;&#x5728;&#x9636;&#x6BB5; 2&#xFF0C;&#x7CA4;&#x6D77;&#x5E02;&#x957F;&#x8FD0;&#x901A;&#x548C;&#x5355;&#x72EC;&#x7684; Xpert PCR &#x4E4B;&#x95F4;&#x7684;&#x534F;&#x8BAE;&#x662F; 95%&#x3002;&#x7B2C; 1 &#x9636;&#x6BB5; Xpert PCR &#x7075;&#x654F;&#x5EA6;&#x662F;&#x9AD8;&#x4E8E;&#x7CA4;&#x6D77; &#xFF08;cyt&#xFF09;&#x3002;NAP1 &#x682A;&#x68C0;&#x6D4B; PCR &#x6838;&#x7CD6;&#x3001; &#x6D4B;&#x5E8F;&#x53CA; Xpert PCR &#x7684;&#x76F8;&#x5173;&#x6027;&#x5F88;&#x597D; (&gt; 90%)&#x3002;&#x4F18;&#x5F02;&#x7684;&#x7075;&#x654F;&#x5EA6;&#x548C;&#x7279;&#x5F02;&#x6027;&#x4EE5;&#x53CA; Xpert pcr &#x4EE5;&#x53CA;&#x5176;&#x5E94;&#x53D8;&#x6253;&#x5B57;&#x80FD;&#x529B;&#x7684;&#x5FEB;&#x901F;&#x5468;&#x8F6C;&#x65F6;&#x95F4;&#x4F7F;&#x5B83;&#x6709;&#x5438;&#x5F15;&#x529B; C.&#x8270;&#x96BE;&#x68AD;&#x83CC;&#x611F;&#x67D3;&#x7684;&#x8BCA;&#x65AD;&#x3002;

Cepheid Xpert &#x68AD;&#x83CC;&#x68AD;&#x72B6;&#x82BD;&#x5B62;&#x6746;&#x83CC;&#x8270;&#x96BE;&#x68AD;&#x83CC;&#x611F;&#x67D3;&#x548C;&#x952E;&#x5165; NAP1 &#x83CC;&#x682A;&#x5728;&#x80BF;&#x7624;&#x4E13;&#x79D1;&#x533B;&#x9662;&#x7684;&#x8BCA;&#x65AD;&#x96BE;&#x8FA8;&#x8BA1;&#x5212;&#x514D;&#x75AB;&#x6D4B;&#x5B9A;&#x65B9;&#x6CD5;&#x7684;&#x8BC4;&#x4EF7;&#x3002;

Clostridium difficile is the most common cause of health care-associated diarrhea. Accurate and rapid diagnosis is essential to improve patient outcome and prevent disease spread. We compared our two-step diagnostic algorithm, an enzyme immunoassay for glutamate dehydrogenase (GDH) followed by the cytotoxin neutralization test (CYT) with a turnaround time of 24 to 48 h, versus the Cepheid Xpert C. difficile Epi assay, a PCR-based assay with a turnaround time of <1 h. In the first phase of the study, only GDH-positive stool samples were tested by both CYT and Xpert PCR. Discordant results were resolved by toxigenic culture. In the second phase, all stool samples were tested by GDH and Xpert PCR. Only GDH-positive stools were further tested by CYT. Genotypic characterization of 45 Xpert PCR-positive stools was performed by sequencing of the tcdC gene and PCR ribotyping. In phase 1, the agreement between the GDH-CYT and the GDH-Xpert PCR was 72%. The sensitivities and specificities of GDH-CYT and GDH-Xpert PCR were 57% and 97% and 100% and 97%, respectively. In phase 2, the agreement between GDH-CYT and Xpert PCR alone was 95%. As in phase 1, sensitivity of the Xpert PCR was higher than that of the GDH-CYT. The correlation between PCR-ribotyping, sequencing, and Xpert PCR for detection of NAP1 strains was excellent (>90%). The excellent sensitivity and specificity and the rapid turnaround time of the Xpert PCR assay as well as its strain-typing capability make it an attractive option for diagnosis of C. difficile infection.

Evaluation of the Cepheid Xpert Clostridium difficile Epi assay for diagnosis of Clostridium difficile infection and typing of the NAP1 strain at a cancer hospital.

Clostridium difficile est la cause la plus fr&eacute;quente de la diarrh&eacute;e associ&eacute;e &agrave; des soins de sant&eacute;. Un diagnostic pr&eacute;cis et rapide est essentiel d&#39;am&eacute;liorer les r&eacute;sultats pour les patients et de pr&eacute;venir la propagation des maladies. Nous avons compar&eacute; notre algorithme diagnostique en deux &eacute;tapes, un test immunoenzymatique de glutamate d&eacute;shydrog&eacute;nase (GDH) suivie de l&#39;&eacute;preuve de s&eacute;roneutralisation cytotoxine (CYT) avec un d&eacute;lai de 24 &agrave; 48 h, contre le test Epi de Cepheid Xpert C. difficile, un test d&#39;amplification PCR dans un d&eacute;lai d&#39;ex&eacute;cution de &lt; 1 h. Dans la premi&egrave;re phase de l&#39;&eacute;tude, seulement les &eacute;chantillons de selles de GDH-positives ont &eacute;t&eacute; test&eacute;s par CYT et Xpert PCR. Les r&eacute;sultats discordants ont &eacute;t&eacute; r&eacute;solues par la culture toxig&eacute;nique. Dans la deuxi&egrave;me phase, tous les &eacute;chantillons de selles ont &eacute;t&eacute; test&eacute;s par GDH et Xpert PCR. Seulement les selles GDH-positives ont &eacute;t&eacute; encore test&eacute;s par CYT. Caract&eacute;risation g&eacute;notypique de 45 tabourets Xpert PCR positif a &eacute;t&eacute; r&eacute;alis&eacute;e par s&eacute;quen&ccedil;age du g&egrave;ne de la CTPD et PCR ribotypage. Dans la phase 1, l&#39;accord entre la GDH-CYT et le PCR de GDH-Xpert &eacute;tait de 72 %. Les sensibilit&eacute;s et les sp&eacute;cificit&eacute;s de GDH-CYT et PCR GDH-Xpert &eacute;taient de 57 % et 97 % et 100 % et 97 %, respectivement. Dans la phase 2, l&#39;accord entre GDH-CYT et Xpert PCR seul &eacute;tait de 95 %. Comme dans la phase 1, sensibilit&eacute; de la PCR Xpert &eacute;tait sup&eacute;rieure &agrave; celui de la GDH-CYT. La corr&eacute;lation entre la PCR-ribotypage, s&eacute;quen&ccedil;age et Xpert PCR pour la d&eacute;tection des souches NAP1 &eacute;tait excellente (&amp;gt; 90 %). L&#39;excellente sensibilit&eacute; et sp&eacute;cificit&eacute; et le d&eacute;lai d&#39;ex&eacute;cution rapide de l&#39;&eacute;preuve de PCR de Xpert ainsi que sa capacit&eacute; d&#39;identification de la souche rendent une option attrayante pour le diagnostic de l&#39;infection &agrave; c. difficile.

Evaluation du test Epi Cepheid Xpert Clostridium difficile pour le diagnostic de l&#39;infection &agrave; Clostridium difficile et le typage de la souche NAP1 dans un h&ocirc;pital du cancer.

Clostridium Difficile ist die h&auml;ufigste Ursache von Gesundheitsf&uuml;rsorge-assoziierter Durchfall. Genaue und schnelle Diagnose ist wichtig zur Verbesserung der Patienten Ergebnis und Verh&uuml;tung von Krankheiten zu verbreiten. Wir verglichen unsere zweistufigen diagnostischen Algorithmus, ein Enzym-Immunoassay f&uuml;r Glutamatdehydrogenase (GDH) gefolgt von der Antigen-Neutralisierung-Test (CYT) mit einer Bearbeitungszeit von 24 bis 48 h, im Vergleich zu den Cepheiden Xpert C.-Difficile-Epi-Assay, eine PCR-gegr&uuml;ndeten Probe mit einer Bearbeitungszeit von &lt; 1H. In der ersten Phase der Studie wurden nur GDH-positiven Stuhlproben von CYT und Xpert PCR getestet. Unzutreffende Ergebnisse wurden durch toxigenen Kultur gel&ouml;st. In der zweiten Phase wurden alle Stuhlproben von GDH und Xpert-PCR getestet. Nur GDH-Positive Hocker wurden weiter von CYT getestet. Genotypische Charakterisierung 45 Xpert PCR-Positive Hocker wurde durch Sequenzierung der TcdC gen und PCR-Ribotyping durchgef&uuml;hrt. In Phase 1 war ein Vertrag zwischen der GDH-CYT und der GDH-Xpert-PCR 72 %. Die Befindlichkeiten und Besonderheiten der GDH-CYT und GDH-Xpert-PCR wurden 57 % und 97 % und 100 % und 97 %. In Phase 2 war ein Vertrag zwischen GDH-CYT und Xpert PCR allein 95 %. Wie in Phase 1 lag die Sensitivit&auml;t des Xpert PCR h&ouml;her als der der GDH-CYT. Die Korrelation zwischen PCR-Ribotyping, Sequenzierung und Xpert PCR zum Nachweis von NAP1 St&auml;mme war ausgezeichnet (&gt; 90 %). Die hervorragende Sensitivit&auml;t und Spezifit&auml;t und die schnelle Bearbeitungszeit von Xpert PCR-Assays sowie seine F&auml;higkeit Stammtypisierung machen es attraktiv f&uuml;r C. Difficile-Infektionen.

Bewertung des Cepheiden Xpert Clostridium Difficile Epi Tests zur Diagnose von Clostridium Difficile Infektion und Eingabe der Belastung NAP1 in einem Krankenhaus Krebs.

Clostridium difficile &egrave; la causa pi&ugrave; comune di diarrea associate all&#39;assistenza sanitaria. Diagnosi rapida e accurata &egrave; essenziale per migliorare il risultato paziente e prevenire le malattie diffuse. Abbiamo confrontato il nostro algoritmo diagnostico in due fasi, saggio immunoenzimatico per la glutammato deidrogenasi (GDH) seguita dal test di neutralizzazione citotossina (CYT) con un tempo di consegna di 24-48 h, versus il dosaggio di Epi Cefeidi Xpert C. difficile, un saggio PCR-based con un tempo di turnaround di &lt; h 1. Nella prima fase dello studio, solo campioni di feci GDH-positivi sono stati testati da CYT sia Xpert PCR. Risultati discordanti sono stati risolti da tossinogenici. Nella seconda fase, tutti i campioni di feci sono stati testati da GDH e Xpert PCR. Solo sgabelli GDH-positivi sono stati ulteriormente testati da CYT. Caratterizzazione genotipica di 45 sgabelli Xpert PCR positiva &egrave; stata eseguita mediante sequenziamento del gene tcdC e PCR Ribotipia. Nella fase 1, l&#39;accordo tra la GDH-CYT e PCR GDH-Xpert era 72%. La sensibilit&agrave; e la specificit&agrave; della GDH-CYT e GDH-Xpert PCR erano 57% e 97% e 100% e 97%, rispettivamente. Nella fase 2, l&#39;accordo tra GDH-CYT e Xpert PCR da solo era il 95%. Come nella fase 1, sensibilit&agrave; della PCR Xpert &egrave; stato superiore a quello della GDH-CYT. La correlazione tra PCR-Ribotipia, sequencing e Xpert PCR per la rilevazione di ceppi NAP1 era eccellente (&amp;gt; 90%). L&#39;eccellente sensibilit&agrave; e specificit&agrave; e il tempo di consegna rapida di dosaggio Xpert PCR, come pure la sua capacit&agrave; tipizzazione lo rendono un&#39;opzione attraente per la diagnosi dell&#39;infezione da c. difficile.

Valutazione del dosaggio Cefeidi Xpert Clostridium difficile Epi per la diagnosi di infezione da Clostridium difficile e la tipizzazione del ceppo NAP1 presso un ospedale di cancro.

&#x30AF;&#x30ED;&#x30B9;&#x30C8;&#x30EA;&#x30B8;&#x30A6;&#x30E0; &#x30FB; &#x30C7;&#x30A3;&#x30D5;&#x30A3;&#x30B7;&#x30EB; &#x30D8;&#x30EB;&#x30B9;&#x30B1;&#x30A2;&#x95A2;&#x9023;&#x4E0B;&#x75E2;&#x306E;&#x6700;&#x3082;&#x4E00;&#x822C;&#x7684;&#x306A;&#x539F;&#x56E0;&#x3067;&#x3059;&#x3002;&#x6B63;&#x78BA;&#x3067;&#x8FC5;&#x901F;&#x306A;&#x8A3A;&#x65AD;&#x306F;&#x60A3;&#x8005;&#x306E;&#x30A2;&#x30A6;&#x30C8;&#x30AB;&#x30E0;&#x3092;&#x6539;&#x5584;&#x3057;&#x3001;&#x75C5;&#x6C17;&#x306E;&#x8513;&#x5EF6;&#x3092;&#x9632;&#x3050;&#x305F;&#x3081;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3059;&#x3002;&#x6211;&#x3005; &#x306F;&#x79C1;&#x305F;&#x3061; 2 &#x6BB5;&#x968E;&#x8A3A;&#x65AD;&#x30A2;&#x30EB;&#x30B4;&#x30EA;&#x30BA;&#x30E0;&#x306B;&#x6BD4;&#x3079;&#x3066;&#x3001;&#x91CE;&#x5916;&#x3088;&#x308A;&#x63A1;&#x53D6;&#x3057;&#x305F;&#x4E2D;&#x548C;&#x8A66;&#x9A13; (CYT) &#x5BFE;&#x30BB;&#x30D5;&#x30A1;&#x30A4;&#x30C9; Xpert c. &#x30C7;&#x30A3;&#x30D5;&#x30A3;&#x30B7;&#x30EB; Epi &#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306E; 24 &#x301C; 48 &#x6642;&#x9593;&#x306E;&#x30BF;&#x30FC;&#x30F3;&#x30A2;&#x30E9;&#x30A6;&#x30F3;&#x30C9;&#x6642;&#x9593;&#x3092;&#x9175;&#x7D20;&#x30A4;&#x30E0;&#x30CE;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306B;&#x3088;&#x308B;&#x30B0;&#x30EB;&#x30BF;&#x30DF;&#x30F3;&#x9178;&#x8131;&#x6C34;&#x7D20;&#x9175;&#x7D20; (GDH) PCR &#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306E;&#x30BF;&#x30FC;&#x30F3;&#x30A2;&#x30E9;&#x30A6;&#x30F3;&#x30C9;&#x6642;&#x9593;&#x7D9A;&#x3044;&#x3066; &lt; 1 h&#x3002;&#x7814;&#x7A76;&#x306E;&#x6700;&#x521D;&#x306E;&#x6BB5;&#x968E;&#x3067;&#x306E;&#x307F; GDH &#x6B63;&#x7CDE;&#x4FBF; CYT &#x3068; Xpert PCR &#x306B;&#x3088;&#x3063;&#x3066;&#x30C6;&#x30B9;&#x30C8;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x8033;&#x969C;&#x308A;&#x306A;&#x7D50;&#x679C;&#x306F;&#x3001;&#x6BD2;&#x7D20;&#x7523;&#x751F;&#x6587;&#x5316;&#x306B;&#x3088;&#x3063;&#x3066;&#x89E3;&#x6C7A;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;2 &#x756A;&#x76EE;&#x306E;&#x30D5;&#x30A7;&#x30FC;&#x30BA;&#x3067;&#x306F;&#x3001;&#x3059;&#x3079;&#x3066;&#x306E;&#x4FBF;&#x306E;&#x30B5;&#x30F3;&#x30D7;&#x30EB; GDH &#x304A;&#x3088;&#x3073; Xpert PCR &#x306B;&#x3088;&#x3063;&#x3066;&#x691C;&#x67FB;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;GDH &#x6B63;&#x30B9;&#x30C4;&#x30FC;&#x30EB;&#x3060;&#x3051;&#x306F;&#x3001;&#x3055;&#x3089;&#x306B; CYT &#x306B;&#x3088;&#x3063;&#x3066;&#x30C6;&#x30B9;&#x30C8;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;TcdC &#x907A;&#x4F1D;&#x5B50;&#x3068; PCR ribotyping &#x306E;&#x914D;&#x5217;&#x306B;&#x3088;&#x3063;&#x3066; 45 Xpert PCR &#x967D;&#x6027;&#x30B9;&#x30C4;&#x30FC;&#x30EB;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x578B;&#x306B;&#x3088;&#x308B;&#x30AD;&#x30E3;&#x30E9;&#x30AF;&#x30BF;&#x30EA;&#x30BC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x884C;&#x3063;&#x305F;&#x3002;&#x30D5;&#x30A7;&#x30FC;&#x30BA; 1 &#x3067;&#x306F;&#x3001;GDH CYT &#x3068; GDH Xpert PCR &#x306E;&#x5408;&#x610F; 72% &#x3067;&#x3042;&#x3063;&#x305F;&#x3002;&#x611F;&#x5EA6;&#x3068;&#x7279;&#x7570;&#x6027;&#x306E; GDH CYT &#x3068; GDH Xpert PCR &#x306F;&#x305D;&#x308C;&#x305E;&#x308C; 57% &#x3068; 97% &#x3068; 100% &#x3068; 97% &#x3067;&#x3042;&#x3063;&#x305F;&#x3002;&#x30D5;&#x30A7;&#x30FC;&#x30BA; 2 &#x3067;&#x306F;&#x3001;GDH CYT &#x3068; Xpert PCR &#x3060;&#x3051;&#x3067;&#x306E;&#x5408;&#x610F;&#x306F; 95 &#xFF05; &#x3060;&#x3063;&#x305F;&#x3002;&#x30D5;&#x30A7;&#x30FC;&#x30BA; 1 &#x3068;&#x540C;&#x69D8;&#x306B;&#x3001;Xpert PCR &#x306E;&#x611F;&#x5EA6;&#x306F; GDH CYT &#x3088;&#x308A;&#x9AD8;&#x3044;&#x3060;&#x3063;&#x305F;&#x3002;NAP1 &#x83CC;&#x682A;&#x306E;&#x691C;&#x51FA;&#x7528; PCR ribotyping&#x3001;&#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x3001;&#x304A;&#x3088;&#x3073; Xpert PCR &#x306E;&#x76F8;&#x95A2;&#x306F;&#x512A;&#x79C0;&#x3060;&#x3063;&#x305F; &#xFF08;&#x76EE; 90%)&#x3002;&#x512A;&#x308C;&#x305F;&#x611F;&#x5EA6;&#x3068;&#x7279;&#x7570;&#x6027;&#x3068;&#x8FC5;&#x901F;&#x306A;&#x30BF;&#x30FC;&#x30F3;&#x30A2;&#x30E9;&#x30A6;&#x30F3;&#x30C9;&#x6642;&#x9593; Xpert PCR &#x6CD5;&#x3068;&#x305D;&#x306E;&#x3072;&#x305A;&#x307F;&#x5165;&#x529B;&#x6A5F;&#x80FD;&#x306E;&#x9B45;&#x529B;&#x7684;&#x306A;&#x30AA;&#x30D7;&#x30B7;&#x30E7;&#x30F3; c. &#x30C7;&#x30A3;&#x30D5;&#x30A3;&#x30B7;&#x30EB;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x8A3A;&#x65AD;&#x3092;&#x4F5C;&#x308B;&#x3002;

&#x30BB;&#x30D5;&#x30A1;&#x30A4;&#x30C9; Xpert &#x30AF;&#x30ED;&#x30B9;&#x30C8;&#x30EA;&#x30B8;&#x30A6;&#x30E0; &#x30FB; &#x30C7;&#x30A3;&#x30D5;&#x30A3;&#x30B7;&#x30EB; Epi &#x30A2;&#x30C3;&#x30BB;&#x30A4; &#x30AF;&#x30ED;&#x30B9;&#x30C8;&#x30EA;&#x30B8;&#x30A6;&#x30E0; &#x30FB; &#x30C7;&#x30A3;&#x30D5;&#x30A3;&#x30B7;&#x30EB;&#x611F;&#x67D3;&#x3068;&#x304C;&#x3093;&#x306E;&#x75C5;&#x9662; NAP1 &#x3072;&#x305A;&#x307F;&#x306E;&#x30BF;&#x30A4;&#x30D4;&#x30F3;&#x30B0;&#x306E;&#x8A3A;&#x65AD;&#x306E;&#x8A55;&#x4FA1;&#x3002;

Clostridium &#xB0A8;&#xACFC; &#xC5B4;&#xC6B8;&#xB9AC;&#xC9C0; &#xC54A;&#xB294; &#xC758;&#xB8CC; &#xAD00;&#xB828; &#xC124;&#xC0AC;&#xC758; &#xAC00;&#xC7A5; &#xD754;&#xD55C; &#xC6D0;&#xC778;&#xC785;&#xB2C8;&#xB2E4;. &#xC815;&#xD655; &#xD558; &#xACE0; &#xC2E0;&#xC18D;&#xD55C; &#xC9C4;&#xB2E8; &#xD658;&#xC790; &#xACB0;&#xACFC; &#xAC1C;&#xC120; &#xD558; &#xACE0; &#xC9C8;&#xBCD1; &#xD655;&#xC0B0; &#xBC29;&#xC9C0;&#xC5D0; &#xD544;&#xC218;&#xC801; &#xC774;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xC6B0;&#xB9AC;&#xC758; 2 &#xB2E8;&#xACC4; &#xC9C4;&#xB2E8; &#xC54C;&#xACE0;&#xB9AC;&#xC998;&#xC5D0; &#xBE44;&#xD574;, &#xC870;&#xBBF8;&#xB8CC; &#xD6A8;&#xC18C; (GDH)&#xC5D0; &#xB300; &#xD55C; &#xD6A8;&#xC18C; immunoassay Cepheid Xpert C. &#xB0A8;&#xACFC; &#xC5B4;&#xC6B8;&#xB9AC;&#xC9C0; &#xC54A;&#xB294; &#xD53C; &#xBD84;&#xC11D; &#xACB0;&#xACFC; &#xB300; 24 ~ 48 h&#xC758; &#xCC98;&#xB9AC; &#xC2DC;&#xAC04;&#xC744; cytotoxin &#xC911;&#xD654; &#xD14C;&#xC2A4;&#xD2B8; (CYT) PCR &#xAE30;&#xBC18; &#xBD84;&#xC11D; &#xACB0;&#xACFC;&#xC758; &#xCC98;&#xB9AC; &#xC2DC;&#xAC04;&#xC744; &#xAC00;&#xC9C4; &#xB4A4; &lt; 1 h. &#xC5F0;&#xAD6C;&#xC758; &#xCCAB; &#xBC88;&#xC9F8; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C;&#xB294; GDH &#xC591;&#xC131; &#xB300;&#xBCC0; &#xC0D8;&#xD50C;&#xB9CC; CYT &#xBC0F; Xpert PCR&#xC5D0; &#xC758;&#xD574; &#xD14C;&#xC2A4;&#xD2B8; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADC0;&#xC5D0; &#xAC70;&#xC2AC;&#xB9AC;&#xB294; &#xACB0;&#xACFC; toxigenic &#xBB38;&#xD654;&#xC5D0; &#xC758;&#xD574; &#xD574;&#xACB0; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB450; &#xBC88;&#xC9F8; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; &#xBAA8;&#xB4E0; &#xB300;&#xBCC0; &#xC0D8;&#xD50C; GDH&#xC640; Xpert PCR&#xC5D0; &#xC758;&#xD574; &#xD14C;&#xC2A4;&#xD2B8; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. GDH &#xC591;&#xC131; &#xBCC0;&#xB9CC; &#xCD94;&#xAC00; CYT&#xC5D0; &#xC758;&#xD574; &#xD14C;&#xC2A4;&#xD2B8;. 45 Xpert PCR &#xC591;&#xC131; &#xBC1C;&#xD310;&#xC758; genotypic &#xD2B9;&#xC131;&#xD654; tcdC &#xC720;&#xC804;&#xC790;&#xC640; PCR ribotyping&#xC758; &#xC5F0;&#xC18D;&#xC5D0; &#xC758;&#xD574; &#xC218;&#xD589; &#xB418;&#xC5C8;&#xB2E4;. 1 &#xB2E8;&#xACC4;&#xC5D0;&#xC11C;&#xB294; GDH CYT GDH Xpert PCR &#xC0AC;&#xC774; &#xACC4;&#xC57D; 72% &#xC774;&#xC5C8;&#xB2E4;. &#xAC10;&#xB3C4; &#xBC0F; &#xD2B9;&#xC774;&#xC131; GDH CYT GDH Xpert PCR&#xC758; 57%&#xC640; 97%&#xC640; 100%&#xC640; 97%&#xB97C; &#xAC01;&#xAC01; &#xAE30;&#xB85D; &#xD588;&#xB2E4;. 2 &#xB2E8;&#xACC4;&#xC5D0;&#xC11C;&#xB294; GDH CYT Xpert PCR &#xD63C;&#xC790; &#xC0AC;&#xC774; &#xACC4;&#xC57D; 95% &#xC774;&#xC5C8;&#xB2E4;. 1 &#xB2E8;&#xACC4; &#xC5D0;&#xC11C;&#xCC98;&#xB7FC; Xpert PCR&#xC758; &#xBBFC;&#xAC10;&#xB3C4; GDH CYT &#xBCF4;&#xB2E4; &#xB192;&#xC558;&#xB2E4;. NAP1 &#xAE34;&#xC7A5;&#xC758; &#xAC80;&#xCD9C;&#xC744; &#xC704;&#xD55C; PCR ribotyping, &#xC2DC;&#xD000;&#xC2F1;, &#xADF8;&#xB9AC;&#xACE0; Xpert PCR&#xC758; &#xC0C1;&#xAD00;&#xAD00;&#xACC4;&#xB294; &#xC6B0;&#xC218; (&amp;gt; 90%). &#xC6B0;&#xC218;&#xD55C; &#xAC10;&#xB3C4; &#xBC0F; &#xD2B9;&#xC774;&#xC131; &#xBC0F; &#xC2A4;&#xD2B8;&#xB808;&#xC778; &#xC785;&#xB825; &#xAE30;&#xB2A5; Xpert PCR &#xBD84;&#xC11D; &#xACB0;&#xACFC;&#xC758; &#xC2E0;&#xC18D;&#xD55C; &#xCC98;&#xB9AC; &#xC2DC;&#xAC04; &#xADF8;&#xAC83; c. &#xAC10;&#xC5FC;&#xC758; &#xC9C4;&#xB2E8; &#xC704;&#xD55C; &#xB9E4;&#xB825;&#xC801;&#xC778; &#xC635;&#xC158;&#xC744; &#xD655;&#xC778;&#xD569;&#xB2C8;&#xB2E4;.

Clostridium &#xAC10;&#xC5FC;&#xACFC; &#xC554; &#xBCD1;&#xC6D0; NAP1 &#xAE34;&#xC7A5;&#xC758; &#xC785;&#xB825;&#xC758; &#xC9C4;&#xB2E8; &#xC704;&#xD55C; Cepheid Xpert Clostridium &#xB0A8;&#xACFC; &#xC5B4;&#xC6B8;&#xB9AC;&#xC9C0; &#xC54A;&#xB294; &#xD53C; &#xBD84;&#xC11D; &#xACB0;&#xACFC;&#xC758; &#xD3C9;&#xAC00;.

Clostridium difficile es la causa m&aacute;s com&uacute;n de diarrea relacionadas con la atenci&oacute;n de la salud. Diagn&oacute;stico exacto y r&aacute;pido es esencial para mejorar el resultado del paciente y evitar la propagaci&oacute;n de la enfermedad. Comparamos nuestro algoritmo diagn&oacute;stico de dos etapas, un ensayo inmunoenzim&aacute;tico de glutamato deshidrogenasa (GDH) seguido por la prueba de neutralizaci&oacute;n de citotoxina (CYT) con un tiempo de respuesta de 24 a 48 h, contra el ensayo de Epi Cepheid Xpert C. difficile, un ensayo basadas en PCR con un tiempo de respuesta de &lt; 1 h. En la primera fase del estudio, s&oacute;lo muestras de heces de GDH-positivos fueron probadas por CYT tanto Xpert PCR. Resultados discordantes se resolvieron mediante cultivo toxig&eacute;nico. En la segunda fase, todas las muestras de heces fueron probadas por GDH y Xpert PCR. Taburetes de GDH-positivo s&oacute;lo se analizaron adem&aacute;s CYT. Caracterizaci&oacute;n genot&iacute;pica de 45 taburetes Xpert PCR positiva fue realizada por la secuencia del gen de la CTPD y PCR ribotyping. En la fase 1, el acuerdo entre la GDH-CYT y la PCR de GDH-Xpert fue 72%. Las sensibilidades y especificidades de GDH-CYT y GDH-Xpert PCR fueron 57% y 97% y 100% y 97%, respectivamente. En la fase 2, el acuerdo entre GDH-CYT y Xpert PCR solo fue 95%. Como en la fase 1, la sensibilidad de la PCR Xpert fue mayor que el de la GDH-CYT. La correlaci&oacute;n entre ribotyping-PCR y secuenciaci&oacute;n Xpert PCR para la detecci&oacute;n de cepas de NAP1 fue excelente (&gt; 90%). La excelente sensibilidad y especificidad y el tiempo de respuesta r&aacute;pida del ensayo de PCR Xpert, as&iacute; como su capacidad de cepas lo hacen una opci&oacute;n atractiva para el diagn&oacute;stico de infecci&oacute;n por C. difficile.

Evaluaci&oacute;n del ensayo Cepheid Xpert Clostridium difficile Epi para diagn&oacute;stico de infecci&oacute;n por Clostridium difficile y la tipificaci&oacute;n de la cepa de NAP1 en un hospital de c&aacute;ncer.

Respiratory viruses are increasingly recognized as serious causes of morbidity and mortality in immunocompromised patients. The rapid and sensitive detection of respiratory viruses is essential for the early diagnosis and administration of appropriate antiviral therapy, as well as for the effective implementation of infection control measures. We compared the performance of two commercial assays, xTAG RVP Fast (Luminex Diagnostics, Toronto, Canada) and FilmArray RVP (FA RVP; Idaho Technology, Salt Lake City, UT), in pediatric patients at Memorial Sloan-Kettering Cancer Center. These assays detect the following viruses: respiratory syncytial virus; influenza A and B viruses; parainfluenza viruses 1, 2, 3, and 4; human metapneumovirus; adenovirus; enterovirus-rhinovirus; coronaviruses NL63, HKU1, 229E, and OC43; and bocavirus. We tested a total of 358 respiratory specimens from 173 pediatric patients previously tested by direct fluorescence assay (DFA) and viral culture. The overall detection rate (number of positive specimens/total specimens) for viruses tested by all methods was 24% for DFA/culture, 45% for xTAG RVP Fast, and 51% for FA RVP. The agreement between the two multiplex assays was 84.5%, and the difference in detection rate was statistically significant (P < 0.0001). Overall, the FA RVP assay was more sensitive than the xTAG RVP Fast assay and had a turnaround time of approximately 1 h. The sensitivity, simplicity, and random-access platform make FA RVP an excellent choice for laboratory on-demand service with low to medium volume.

Comparison of the Luminex xTAG RVP Fast assay and the Idaho Technology FilmArray RP assay for detection of respiratory viruses in pediatric patients at a cancer hospital.

We describe here the use of an immunomagnetic separation enrichment process coupled with a modified real-time cellular analysis (RTCA) system (RTCA version 2) for the detection of C. difficile toxin (CDT) in stool. The limit of CDT detection by RTCA version 2 was 0.12 ng/ml. Among the consecutively collected 401 diarrheal stool specimens, 53 (13.2%) were toxin-producing C. difficile strains by quantitative toxigenic culture (qTC); bacterial loads ranged from 3.00 × 10(1) to 3.69 × 10(6) CFU/ml. The RTCA version 2 method detected CDT in 51 samples, resulting in a sensitivity of 96.2%, a specificity of 99.7%, and positive and negative predictive values of 98.1% and 99.4%, respectively. The positive step time ranged from 1.43 to 35.85 h, with <24 h for 80% of the samples. The CDT concentrations in stool samples determined by RTCA version 2 correlated with toxigenic C. difficile bacterial load (R(2) = 0.554, P = 0.00002) by qTC as well as the threshold cycle (R(2) = 0.343, P = 0.014) by real-time PCR. A statistically significant correlation between the CDT concentrations and the clinical severity of CDI was observed (P = 0.015). The sensitivity of the RTCA version 2 assay for the detection of functional toxins in stool specimens was significantly improved when the immunomagnetic separation enrichment process was incorporated. More than 80% positive results can be obtained within 24 h. The stool specimen CDT concentration derived using the RTCA version 2 assay correlates with clinical severity and may be used as a marker for monitoring the status of CDI.

Real-time cellular analysis coupled with a specimen enrichment accurately detects and quantifies Clostridium difficile toxins in stool.

Rapid and accurate diagnosis of influenza is important for infection control, as well as for patient management. Alere i Influenza A&B is an isothermal nucleic acid amplification-based integrated system for detection and differentiation of influenza virus A and influenza virus B. The performance of the Alere i Influenza A&B was screened using frozen nasopharyngeal-swab specimens collected in viral transport medium (VTM) that were originally tested fresh with the FilmArray Respiratory Panel (RP) assay during the 2012-2013 influenza outbreak. In total, 360 VTM specimens were selected for Alere i Influenza A&B testing: 40 influenza virus A H1N1-2009 (influenza virus A-1), 40 influenza virus A H3N2 (influenza virus A-3), 37 influenza virus A "equivocal" or "no subtype detected" (influenza virus A-u), 41 influenza virus B, and 202 influenza virus-negative specimens, as initially determined by the FilmArray RP assay. The Alere assay showed sensitivities of 87.2%, 92.5%, 25.0%, and 97.4% for influenza virus A-1, influenza virus A-3, influenza virus A-u, and influenza virus B, respectively, after discordant resolution by Prodesse ProFLU+ PCR. The specificities were 100% for both influenza virus A and influenza virus B. In general, the Alere i Influenza A&B provided good sensitivity, although the assay did show poorer sensitivity with samples determined to have low influenza virus A titers by Prodesse ProFlu+ PCR (a mean real-time PCR threshold cycle [CT] value of 31.9 ± 2.0), which included the majority of the samples called influenza virus A "equivocal" or "no subtype detected" by a single BioFire FilmArray RP test. The integrated, rapid, and simple characteristics of the Alere i Influenza A&B assay make it a potential candidate for point-of-care testing, with a test turnaround time of less than 15 min.

Evaluation of Alere i Influenza A&B for rapid detection of influenza viruses A and B.

We evaluated the Lyra Direct HSV 1+2/VZV multiplex real-time PCR assay for the detection and differentiation of herpes simplex virus 1 (HSV-1), HSV-2, and varicella-zoster virus (VZV) on 695 consecutive cutaneous and mucocutaneous lesion specimens. The intra-assay and interassay coefficient of variation values for the Lyra assay were 0.29 to 1.30% and 2.33 to 2.61%, respectively. The sensitivities, specificities, and positive and negative predictive values were 93.4 to 95.0%, 96.1 to 96.8%, 78.0 to 80.3%, and 99.0 to 99.1%, respectively, in comparison to those of viral culture. The values were further improved when a resolution analysis was performed with a laboratory-developed PCR assay.

Clinical validation of the Lyra direct HSV 1+2/VZV assay for simultaneous detection and differentiation of three herpesviruses in cutaneous and mucocutaneous lesions.

Cytomegalovirus (CMV) viral loads in hematopoietic stem cell transplant (HSCT) recipients are typically monitored using quantitative molecular assays. The Roche Cobas AmpliPrep/Cobas TaqMan CMV test (Cobas CMV) has recently been cleared by the FDA for the monitoring of CMV viral loads in plasma samples from transplant patients. In this study, we compare and correlate the viral loads obtained by a laboratory-developed test (LC CMV) (using Roche analyte-specific reagents [ASR] on the LightCycler 2.0) on whole-blood specimens with those obtained on corresponding plasma and whole-blood specimens by the Cobas CMV assay. Testing was performed on 773 archived patient specimens. The strength of the agreement was good for the two assays performed on whole blood (κ=0.6; 95% confidence interval [CI], 0.51 to 0.7) and moderate when the tests were performed on different sample types (κ=0.54; 95% CI, 0.47 to 0.62 for the LC CMV whole blood [WB] assay versus Cobas plasma [PL], and κ=0.57; 95% CI, 0.5 to 0.65 for the Cobas WB assay versus Cobas PL), although the difference was not statistically significant. Using a combination gold standard (i.e., a true positive was a specimen that was positive by two or more methods), the sensitivity and specificity of the assays were 78.8% and 99.3% for the LC CMV assay, 85.2% and 98.1% for the Cobas CMV WB assay, and 100% and 90.5% for Cobas CMV PL assay, respectively. A comparison of the CMV viral load trends in both plasma and whole blood from a few patients with multiple positive successive samples showed similar slopes, with differences in the slope ranging from 0.01 to 0.22. However, the absolute value for individual viral load differed markedly with whole-blood viral loads, being on average 0.5- to 1.22-log higher than those in plasma. The Cobas CMV assay provides a valid option for the monitoring of viral loads in transplant patients. Due to its increased sensitivity, the detection of CMV DNA in patients with low viral loads (i.e., those below limit of quantification [LOQ]) is increased with the Cobas CMV assay in plasma specimens. Longitudinal prospective studies will be needed to examine the clinical significance of these low-level viral loads.

Monitoring of cytomegalovirus viral loads by two molecular assays in whole-blood and plasma samples from hematopoietic stem cell transplant recipients.

Macrolide-resistant Mycoplasma pneumoniae (MRMP) is highly prevalent in Asia and is now being reported from Europe. Few data on MRMP are available in the United States. Using genotypic and phenotypic methods, we detected high-level MRMP in 13.2% of 91 M. pneumoniae--positive specimens from 6 US locations.

Macrolide-Resistant Mycoplasma pneumoniae, United States.

An evaluation of the Luminex NxTAG Respiratory Pathogen Panel was performed on 404 clinical respiratory specimens. Clinical sensitivities and specificities of the assay compared to those of the reference methods were 80.0% to 100.0% and 98.9% to 100.0%, respectively. Correct genotyping information was provided for 95.5% of influenza virus A specimens. The closed-tube format of the assay simplified the workflow and minimized carryover contamination.

Jeffrey Stiles

Department of Laboratory Medicine

Memorial Sloan Kettering Cancer Center

Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Jeffrey Stiles

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Laboratory Medicine

Memorial Sloan Kettering Cancer Center

Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Department of Laboratory Medicine