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DIAGNOSTIK KUMAN MYCOBACTERIUM TUBERCULOSIS . Diagnosis Kuman... diagnostik kuman mycobacterium tuberculosis dengan teknik zhiel neelsen & pcr (polymerase chain reaction) serta resistensinya

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Text of DIAGNOSTIK KUMAN MYCOBACTERIUM TUBERCULOSIS . Diagnosis Kuman... diagnostik kuman mycobacterium...

  • DIAGNOSTIK KUMAN MYCOBACTERIUM TUBERCULOSIS DENGAN

    TEKNIK ZHIEL NEELSEN & PCR (POLYMERASE CHAIN REACTION)

    SERTA RESISTENSINYA DENGAN TEKNIK GENEXPERT MTB/RIF

    DR. dr. Mukhtar Ikhsan, MARS,SpP(K), Yuliati, MBiomed, Novi Prasetyowati, Ssi

    FKIK UIN Syarif Hidayatullah Jakarta, DR.dr.Mukhtar Ikhsan, MARS, SpP(K)

    [email protected]

    FKIK UIN Syarif Hidayatullah Jakarta, Yuliati, MBiomed

    [email protected]

    FKIK UIN Syarif Hidayatullah Jakarta, Novi Prasetyowati

    [email protected]

    ABSTRACT

    Diagnostic of Mycobacterium tuberculosis with Technique of Zhiel Neelsen & PCR (Polymerase Chain Reaction) and Resistance by MTB / RIF Genexpert Technique, 2017.

    Tuberculosis (TB) is the most common cause of death in developing countries of which 75% of patients are in the productive age of 20-49 years, because in developing countries have dense population and high prevalence, so more than 65% cases of tuberculosis occur in Asia.Terjangsung increase in this case is caused by the immune system, nutritional status and personal hygiene of individuals and density of residential neighborhood. WHO recommends treatment for Pulmonary TB patients with DOTS strategy (Directly Observed Treatment Shortcourse Chemotherapy) -or direct supervision of short-term and daily medication-only 36% with an 87% cure rate. Due to the irregular treatment and inadequate combination of drugs in the past there may be immunity of TB germs against widespread or multi drug drug resistance (MDR). This study aims to determine the resistance and sensitivity of Mycobacterium tuberculosis to Anti Tuberculosis drug in tuberculosis patients with PCR and GeneXpert technique. The research used laboratory experimental method which was conducted in laboratory of Microbiology Faculty of Medicine and Health Sciences of Syarif Hidayatullah State Islamic University Jakarta in May 2017. The method used was identification of bacteria with Zhiel Neelsen staining, Homogenase and Decontaminase, Biochemical Test of Mycobacterium tuberculosis, PCR, and GeneXpertMTB / RIF. Sputum sampling is done in the morning - while (SPS). The result of BTA staining from Bojonggede Puskesmas was 84 samples, BTA positive was 35 samples and the percentage of BTA positive was 42%, while BTA negative was 49 and the percentage was 58%. PCR test results from 20 samples was 14 positive samples that mean people with tuberculosis infection 70%, while the negative is 6 samples means people who do not suffer from tuberculosis by 30%. The conclusion of this research is PCR test result compared with result of BTA test with Zhiel Neelsen, that is the result of positive test of the percentage is bigger than smear staining test result.

    Keywords: Mycobacterium tuberculosis, mutation, multi drug resistance (MDR), decontamination.

    mailto:[email protected] mailto:[email protected] mailto:[email protected]

  • COMPARISON OF DIAGNOSTIC TESTS BACTERI MYCOBACTERIUM

    TUBERCULOSIS WITH STAINING BACTERI RESISTANT ACID ZHIEL

    NEELSEN METHOD AND PCR (POLYMERASE CHAIN REACTION)

    Mukhtar Ikhsan, Yuliati

    INTRODUCTION

    A. Background

    Tuberculosis (TB) is an infectious disease caused by Mycobacterium

    tuberculosis and was discovered over 100 years ago. Until now tuberculosis is still a

    serious problem worldwide, as it is the leading cause of death. Every year an estimated

    8 million new infections occur and 2.5 to 3 million cause death (Zhang et al., 2005). Of

    all cases of tuberculosis in the world, most occur in developing countries, including

    Indonesia is ranked third after the state of India and China, with the number of new

    cases about 539,000 cases and the number of deaths about 101,000 people per year.

    According to the Ministry of Health (2008), the disease is most prevalent in developing

    countries of about 75% of patients are at productive age of 20-49 years, this is because

    in developing countries have a dense population and a high prevalence, resulting in

    more than 65% of cases Tuberculosis occurs in Asia. Based on Household Health

    Survey (SKRT) in 1992, pulmonary tuberculosis in Indonesia was ranked second after

    cardiovascular disease.

    According to WHO (2010) states that Indonesia every year there are 583 new

    cases with the death of 130 patients with positive tuberculosis on dahaknya.Hasil

    research Kusnindar (1990) the number of deaths caused by tuberculosis estimated

    105.952 people per year. The incidence of high pulmonary tuberculosis cases is most

  • prevalent in socioeconomic groups. The increase in cases is influenced by immune

    system, nutritional status and personal hygiene of individuals and density of residential

    neighborhood.

    In 2011 WHO recommends treatment for Pulmonary TB patients with DOTS

    strategy (Directly Observed Treatment Shortcourse Chemotherapy) or direct daily

    swallowing of short-term medication only reaches 36% with an 87% cure rate. Prior to

    the DOTS strategy (1969-1994) the coverage was 56% with a cure rate of only 40-60%.

    Because irregular treatment and inadequate drug combinations in the past may have

    arisen TB germs against OAT (anti-tuberculosis drugs) widespread or multi drug

    resistance (MDR).

    Conventional laboratory tests for TB diagnosis and drug sensitivity testing to

    detect first-line and second-line drug resistance take a long time to know the results and

    with more complicated techniques (Boehme, 2009; WHO, 2013). While the patient is

    waiting for the diagnosis, the patient's illness worsens and the patient can move the

    OAT resistant tuberculosis disease to another, especially to the family member. Early

    case detection of MDR TB is important to inhibit transmission and to prevent further

    spread of MDR TB so that new diagnostic tests are needed (Boehme, 2009).

    Conventional tests include the method of seed culture culture, aimed to

    multiply the bacteria Mycobacterium tuberculosis in sputum specimens, so as to

    improve the detection of sensitivity. Currently there are several media that can be used

    as cultures from Mycobacterium tuberculosis, such as solid and liquid media, such as

    Lowenstein-Jensen, Mycobacteria Growth Indicator Tube (MGIT) and microcolonial

    culture. But all the above checks cost not cheap. Thus, other techniques in kultr research

    are still developing to detect Mycobacterium tuberculosis in order to obtain cheap

  • culture method and high sensitivity and specificity. A diagnostic test of the culture of

    the medium taken to identify the Mycobacterium tuberculosis bacteria has some

    disadvantages including it takes longer to get the results, although the cost can be

    cheaper (Indah W, 2010).

    Conventional tests include the method of seed culture culture, aimed to multiply

    the bacteria Mycobacterium tuberculosis in sputum specimens, so as to improve the

    detection of sensitivity. Currently there are several media that can be used as cultures

    from Mycobacterium tuberculosis, such as solid and liquid media, such as Lowenstein-

    Jensen, Mycobacteria Growth Indicator Tube (MGIT) and microcolonial culture. But

    all the above checks cost not cheap. Thus, other techniques in kultr research are still

    developing to detect Mycobacterium tuberculosis in order to obtain cheap culture

    method and high sensitivity and specificity. A diagnostic test of the culture of the

    medium taken to identify the Mycobacterium tuberculosis bacteria has some

    disadvantages including it takes longer to get the results, although the cost can be

    cheaper (Indah W, 2010).

    According to WHO (2011 & 2010) states that only a small portion of the

    440,000 MDR-TB sufferers have adequately tested drug sensitivity. The most

    important problem is that conventional diagnostic methods are currently slow and

    complicated (O.Grady et al., 2011; WHO, 2008). First required isolation of

    Mycobacterium tuberculosis (M. tuberculosis) on the specimen, then identified, and

    continued with the OAT sensitivity test. The test of this recommended World Health

    Organization (WHO) is a method of proportion using media such as Lowenstein Jensen,

    which takes 8- 12 weeks. During that time the patient received an inappropriate drug

    and an increase in the number of M. mutant tuberculosis (WHO, 2008 & Chiang CY,

    2010).

  • According to Diana et al (2010) has conducted research with a diagnostic test

    designed by cross sectional. Sputum examination of patients is done by three

    examination techniques, namely by PCR techniques, examination of BTA

    microscopically, and bacterial culture. The results of the study by comparing the

    method of examination of Acid Resistant Bacteria microskospik with PCR technique

    to detect Mycobacterium tuberculosis has a sensitivity of 30%, specificity 80%, and

    accuracy of 47%. The significance test with Mc Nemar yielded significant differences.

    (p

  • mycobacteria isolate. The M. tuberculosis germ on 21 sputum samples from 30 samples

    consisting of 25 samples of AFB + and 5 smear samples -, can be detected with nested

    PCR.Nested PCR using primers designed from the rpo ~ M. tuberculosis gene section

    is a fast, sen

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