Jornada de Genotipagem

Jornada de Genotipagem
TRUGENE HIV-1 Chamados: Setembro 2011 a Agosto 2012 Jornada de Genotipagem 2012

Objetivos Análise Estatística dos chamados Principais chamados

Fluxo de Atendimento - 0800 770 33 99 0800 770 33 99 N S
Cliente liga para CSC Chamada é tratada por um Técnico ou Assessor Científico Problema Solucionado? Chamado é registrado em sistema e finalizado Chamado é agendado para visita. Representante executa o serviço, e registra as informações em sistema. S N Tenha sempre o número de série ou ativo em mãos Solicite o número do chamado

Pontos de Atenção Todo e qualquer problema ou dúvida deve ser registrado via abertura de chamado através do O envio de mensagens para o NÃO exclui a necessidade de abertura de chamado. Copyright © Siemens AG All rights reserved.

Setembro- 2011 a Agosto- 2012 – TOTAL: 246 Chamados

Visitas

Distribuição Mensal de Chamados

Classificação dos Chamados

Classificação Motivos – Hardware

Pontos de Atenção - Hardware
Cross Talk Gel Toaster Buffer Chamber + Current Leak Controle de Temperatura

Cross-talk Desvios da Linha de Base causados por sombra entre as leituras dos canais Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. TRUGENE HIV-1 Training Program for IVD Use

VERIFICAÇÃO DO SISTEMA ÓPTICO (T-TRACK )
Cy5 bandpass Cy5.5 bandpass Cy5.5 dye spectra Dichroic response Cy5 dye spectra Cy5.5 ‘crosstalk’ into 5 channel Cy5 ‘crosstalk’ into 5.5 channel Wavelength Amplitude Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. TRUGENE HIV-1 Training Program for IVD Use

SINTOMAS DE CROSSTALK ACIMA DE 5%
A análise da corrida e verificação pode ser feita no Multi Viewer. Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. TRUGENE HIV-1 Training Program for IVD Use

Observar o alinhamento da linha base dos canais 5’ e 3’. Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. TRUGENE HIV-1 Training Program for IVD Use

No Raw Data Viewer, selecionar uma mesma região para os dois canais ( pico ) e fazer o cálculo. 164÷1697x100=9,66% Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. TRUGENE HIV-1 Training Program for IVD Use

Corrida com CrossTalk OK ( abaixo de 5% ) Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. 19 ÷ 3246 x 100 = 0,58% TRUGENE HIV-1 Training Program for IVD Use

Corrida com CrossTalk FORA DOS PADRÕES ( Acima de 5% ) Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. 215 ÷ 2742 x 100 = 7,84% TRUGENE HIV-1 Training Program for IVD Use

GEL TOASTER – PRESILHA E BORRACHA DE VEDAÇÃO
1 – Observe se há folga na presilha de travamento da placa de vidro Microcel no toaster. 2 – Observe se há uma deformidade na borracha do fill fixture, ocasionando vazamento do Surefill.

ERROR 6 – Current leak CHECK MGC
1 – Observar se as travas da câmara de tampão estão travando completamente a mesma. 2 – Verificar o volume da solução de tampão 3 – Verificar a situação dos eletrodos

Controle de temperatura
SIL PAD TRAVAS Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. 1 – Observar se as travas da câmara de tampão estão travando completamente a mesma. 2 – Verificar se o Sil Pad não esta danificado. TRUGENE HIV-1 Training Program for IVD Use

Classificação Motivos – Software

Pontos de Atenção - Software
Perda de Comunicação Backup e Indexação dos Dados

CONEXÃO ENTRE TORRE E COMPUTADOR
Perda de Comunicação CONEXÃO ENTRE TORRE E COMPUTADOR Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. Verificar se o cabo do Keyspan ( cinza ) se encontra bem encaixado entre o mesmo e o computador ( entrada USB ). TRUGENE HIV-1 Training Program for IVD Use

CONEXÃO ENTRE TORRE E COMPUTADOR
Led Vermelho Portas Seriais e Led Verde OK OK Positive Control: 1.The positive control comes in 5 single-use tubes that are included in the kit box. 2.Use one of the single-use tubes of positive control, then discard the tube. Do not attempt to refreeze aliquots of positive control for later use. 3. The positive control does not require extraction. It is already extracted wheat germ RNA that has the sequence of the HIV-B1 library. 4. A positive control must be added to each plate if doing groups of 4 samples one tray at a time. If doing batches, follow the recommendations listed in the later slides that are specific to batching. 5. The positive control must be analyzed and a case created so that it can be used for Genetic Fingerprinting. Negative Control: The negative control comes in 1 single use tube and is stored in the TRUGENE kit box at –20  C with the other kit components. 2.The maximum number of freeze-thaws recommended is 4 after the initial use of the kit. This is possible if you follow the instructions to only set up full plates of samples each time. 3. The negative control MUST be run on every plate to check for contamination. 4. The negative control does not require RNA extraction. It is a wheat germ RNA. The positive and negative controls are always placed at the end of the reaction tray behind the patient samples. They are always placed on the same row so as to control for cross contamination. The positive always follows the patient samples and the negative is always last on the row. Verificar se o Led do Keyspan ( vermelho ) se encontra aceso, indicando que o mesmo está conectado e comunicando com o computador. Junto das entradas Seriais ( do Keyspan )se encontram Leds verdes que indicam que a Torre está Comunicando com aquela entrada do Keyspan. TRUGENE HIV-1 Training Program for IVD Use

Backup e Indexação de Dados

Copiando ou Transferindo arquivos para o backup database

Alerta de Transferência ou Cópia para o backup file e janela Transcript

Gravando o backup database em CD/DVD

Classificação Motivos – Periféricos

Pontos de Atenção - Periféricos
Impressora Estabilizador/No-Break Multicanal Expansível

Classificação Motivos – Consumíveis

PONTOS DE ATENÇÃO – Assessoria Científica

Critérios de Avaliação dos Ensaios
Referência: Manual Operacional RENAGENO – Página 37

Critérios de Avaliação dos Cases

Suspeita de Contaminação

Edição de Inserções e Deleções
Inserções conhecidas na protease: Protease: região dos códons de 35 a 40 E35EG - Paolucci et. al. Antiviral Res 2006; 69: Kozisek et. al. J Virol 2008; 82: E35ETN - Paolucci et. al. Antiviral Res 2006; 69: M36TNL - Grant et. al. Antivir Ther 2001; 6:44. G40GK - Jordan et al. AIDS Res Hum Retroviruses 2009; 25: Bases inseridas: ACA AAT TTG Códons: T N L Referência: Manual Operacional RENAGENO – Página 44

Edição de Inserções e Deleções
Inserções conhecidas na região do códon RT69: T69SX / T69TX: Rakik et al., J Acq Immune def Syndr 1999; 22: van der Hoek at. al. J Virol 2005; 79: T69SXX / T69TXX: De Antoni et al., J Infect Dis 1997; 176: Tamalet et al., AIDS 1998; 12:F161-F166. Winters et al., J Clin Invest 1998; 102: De Jong et al., AIDS 1999; 13: Ross et al., J Human Virol 1999; 2: Sugiura et al., J Human Virol 1999; 2: Briones et al., Virus Res 2000; 66: Menéndez-Airas et. al. Curr Pharm Design 2006; 12: T69TTRVMG : Lobato et. al. AIDS 2002; 18: T69TIKKKNSE: Harrigan et. al. AIDS Res Hum Retroviruses 2007; 23: T69TSTGKKDST: van der Hoek at. al. J Virol 2005; 79: Bases inseridas: AGT AGT GGT Códons: S S G Referência: Manual Operacional RENAGENO – Página 44

Representação dos Aminoácidos
Quando existirem bases heterozigóticas cujos membros codifiquem o mesmo aminoácido, o aminoácido ainda é apresentado da mesma maneira que na selvagem. Quando existir mais do que um aminoácido possível, são apresentados cardinais (###). Uma exceção ocorre quando há apenas dois aminoácidos possíveis e um deles corresponde ao tipo selvagem; neste caso, o tipo selvagem é subtraído e o outro aminoácido resultante é apresentado em letras maiúsculas. Quando não for possível determinar um aminoácido, por exemplo, quando há uma inserção ou deleção, ou quando o Códon acaba antes do aminoácido estar completo, são apresentados vários pontos de interrogação. Se as bases que formam o códon resultarem em um códon de terminação (Stop Códon), ele será representado como TER. Referência: Manual Operacional RENAGENO – Página 41

Chamada de Bases Manual
A chamada de bases inicial mostra um desalinhamento dos picos de primer e de produto. Com uma nova chamada de bases, eles são realinhados e a qualidade do ensaio melhora significativamente. Referência: Manual Operacional RENAGENO – Página 39

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