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This organism causes tuberculosis. Worldwide, M. tuberculosis causes more deaths than any other single microbial agent. Approximately one-third of the world’s population is infected with this organism. Each year, it is estimated that 1.7 million people die of tuberculosis and that 9 million new cases occur. An estimated 500,000 people are infected with a multidrug-resistant strain of M. tuberculosis.
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M. tuberculosis grows slowly (i.e., it has a doubling time of 18 hours, in contrast to most bacteria, which can double in number in 1 hour or less). Because growth is so slow, cultures of clinical specimens must be held for 6 to 8 weeks before being recorded as negative. M. tuberculosis can be cultured on bacteriologic media, whereas M. leprae cannot. Media used for its growth (e.g., Löwenstein-Jensen medium) contain complex nutrients (e.g., egg yolk) and dyes (e.g., malachite green). The dyes inhibit the unwanted normal flora present in sputum samples.
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M. tuberculosis is an obligate aerobe; this explains its predilection for causing disease in highly oxygenated tissues such as the upper lobe of the lung and the kidney. The acid-fast property of M. tuberculosis (and other mycobacteria) is attributed to long-chain (C78–C90) fatty acids called mycolic acids in the cell wall.
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Cord factor (trehalose dimycolate) is correlated with virulence of the organism. Virulent strains grow in a characteristic “serpentine” cordlike pattern, whereas avirulent strains do not. The organism also contains several proteins, which, when combined with waxes, elicit delayed hypersensitivity. These proteins are the antigens in the purified protein derivative (PPD) skin test (also known as the tuberculin skin test). A lipid located in the bacterial cell wall called phthiocerol dimycocerosate is required for pathogenesis in the lung.
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M. tuberculosis is relatively resistant to acids and alkalis. NaOH is used to concentrate clinical specimens; it destroys unwanted bacteria, human cells, and mucus but not the organism. M. tuberculosis is resistant to dehydration and therefore survives in dried expectorated sputum; this property may be important in its transmission by aerosol.
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Strains of M. tuberculosis resistant to the main antimycobacterial drug, isoniazid (isonicotinic acid hydrazide, INH), as well as strains resistant to multiple antibiotics (called multidrug-resistant or MDR strains), have become a worldwide problem. This resistance is attributed to one or more chromosomal mutations, because no plasmids have been found in this organism. One of these mutations is in a gene for mycolic acid synthesis, and another is in a gene for catalase-peroxidase, an enzyme required to activate INH within the bacterium.
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Transmission & Epidemiology
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M. tuberculosis is transmitted from person to person by respiratory aerosols produced by coughing. The source of the organism is a cavity in the lung that has eroded into a bronchus. The portal of entry is the respiratory tract, and the initial site of infection is the lung. In tissue, it resides chiefly within reticuloendothelial cells (e.g., macrophages). Macrophages kill most, but not all, of the infecting organisms. The ones that survive can continue to infect other adjacent cells or can disseminate to other organs.
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Humans are the natural reservoir of M. tuberculosis. Although some animals, such as cattle, can be infected, they are not the main reservoir for human infection. Most transmission occurs by aerosols generated by the coughing of “smear-positive” people (i.e., those whose sputum contains detectable bacilli in the acid-fast stain). However, about 20% of people are infected by aerosols produced by the coughing of “smear-negative” people.
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In the United States, tuberculosis is almost exclusively a human disease. In developing countries, Mycobacterium bovis also causes tuberculosis in humans. M. bovis is found in cow’s milk, which, unless pasteurized, can cause gastrointestinal tuberculosis in humans.
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The disease tuberculosis occurs in only a small number of infected individuals. In the United States, most cases of tuberculosis are associated with reactivation in elderly, malnourished men. The risk of infection and disease is highest among socioeconomically disadvantaged people, who have poor housing and poor nutrition. These factors, rather than genetic ones, probably account for the high rate of infection among Native Americans, African Americans, and Native Alaskans.
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In the United States, there are approximately 15 million people with latent tuberculosis and 10,000 cases of active disease. Most cases of active disease in the United States are caused by reactivation of latent infection. The risk factors for infection and reactivation (progression) to disease are listed in Table 21–3.
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An overall scheme of pathogenesis by M. tuberculosis is shown in Figure 21–2. It describes primary tuberculosis, which typically results in a Ghon focus in the lower lung. Primary tuberculosis can heal by fibrosis, can lead to progressive lung disease, can cause bacteremia and miliary tuberculosis, or can cause hematogenous dissemination resulting in no immediate disease but with the risk of reactivation in later life.
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If the primary infection heals without causing disease, it is called a latent infection. Of those exposed to M. tuberculosis, approximately 90% develop latent infection and approximately 10% develop disease. Of those who have latent infection, approximately 10% progress to active disease (reactivation) at a later time, whereas 90% remain latent.
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Figure 21–2 also describes secondary tuberculosis with a cavity in the upper lobes. This can cause disease directly or result in reactivation disease in later life with central nervous system lesions, vertebral osteomyelitis (Pott’s disease), or involvement of other organs.
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M. tuberculosis produces no well-recognized exotoxins and does not contain endotoxin in its cell wall. However, M. tuberculosis produces two proteins that appear to play a role in pathogenesis. One is tuberculosis necrotizing toxin (TNT), which cleaves nicotinamide adenine dinucleotide (NAD) within macrophages resulting in death of the infected macrophage. The other is early secreted antigen-6 (ESAT-6), a protein that reduces the innate immune response by reducing gamma interferon production, thereby enhancing the virulence of the organism. The precise role of these proteins in pathogenesis remains to be determined.
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The organism preferentially infects macrophages and other reticuloendothelial cells. M. tuberculosis survives and multiplies within a cellular vacuole called a phagosome. The organism produces a protein called exported repetitive protein that prevents the phagosome from fusing with the lysosome, thereby allowing the organism to escape the degradative enzymes in the lysosome.
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Lesions are dependent on the presence of the organism and the host response. There are two types of lesions:
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Exudative lesions, which consist of an acute inflammatory response and occur chiefly in the lungs at the initial site of infection.
Granulomatous lesions, which consist of a central area of giant cells containing tubercle bacilli surrounded by a zone of epithelioid cells. These giant cells, called Langhans’ giant cells, are an important pathologic finding in tuberculous lesions. A tubercle is a granuloma surrounded by fibrous tissue that has undergone central caseation necrosis. Tubercles heal by fibrosis and calcification.
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The primary lesion of tuberculosis usually occurs in the lungs. The parenchymal exudative lesion and the draining lymph nodes together are called a Ghon complex. Primary lesions usually occur in the lower lobes, whereas reactivation lesions usually occur in the apices. Reactivation lesions also occur in other well-oxygenated sites such as the kidneys, brain, and bone. Reactivation is seen primarily in immunocompromised or debilitated patients.
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Spread of the organism within the body occurs by two mechanisms:
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A tubercle can erode into a bronchus, empty its caseous contents, and thereby spread the organism to other parts of the lungs, to the gastrointestinal tract if swallowed, and to other persons if expectorated.
It can disseminate via the bloodstream to many internal organs. Dissemination can occur at an early stage if cell-mediated immunity fails to contain the initial infection or at a late stage if a person becomes immunocompromised.
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Immunity & Hypersensitivity
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After recovery from the primary infection, resistance to the organism is mediated by cellular immunity (i.e., by CD4-positive T cells and macrophages). The CD4-positive T cells are Th-1 helper T cells (see Chapter 58).
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Circulating antibodies also form, but they play no role in resistance and are not used for diagnostic purposes. Patients deficient in cellular immunity, such as patients with acquired immunodeficiency syndrome (AIDS), are at much higher risk for disseminated, life-threatening tuberculosis. Mutations in the interferon-γ receptor gene are another cause of defective cellular immunity that predisposes to severe tuberculosis. This emphasizes the importance of activation of macrophages by interferon-γ in the host defense against M. tuberculosis.
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Prior infection can be detected by a positive tuberculin skin test result, which is due to a delayed hypersensitivity reaction. PPD is used as the antigen in the tuberculin skin test. The intermediate-strength preparation of PPD, which contains five tuberculin units, is usually used. The skin test is evaluated by measuring the diameter of the induration surrounding the skin test site (Figure 21–3). Note that induration (thickening), not simply erythema (reddening), must be observed.
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The diameter required to judge the test as positive varies depending on the status of the individual being tested. Induration of 15 mm or more is positive in a person who has no known risk factors. Induration of 10 mm or more is positive in a person with high-risk factors, such as a homeless person, an intravenous drug user, or a nursing home resident. Induration of 5 mm or more is positive in a person who has deficient cell-mediated immunity (e.g., AIDS patients) or has been in close contact with a person with active tuberculosis.
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A positive skin test result indicates previous infection by the organism but not necessarily active disease. The tuberculin test becomes positive 4 to 6 weeks after infection. Immunization with bacillus Calmette-Guérin (BCG) vaccine (see page 185) can cause a positive test, but the reactions are usually only 5 to 10 mm and tend to decrease with time. People with PPD reactions of 15 mm or more are assumed to be infected with M. tuberculosis even if they have received the BCG vaccine. A positive skin test reverts to negative in about 5% to 10% of people. Reversion to negative is more common in the United States now than many years ago because now a person is less likely to be exposed to the organism and therefore less likely to receive a boost to the immune system.
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The skin test itself does not induce a positive response in a person who has not been exposed to the organism. It can, however, “boost” a weak or negative response in a person who has been exposed to produce a positive reaction. The clinical implications of this “booster effect” are beyond the scope of this book.
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Tuberculin reactivity is mediated by the cellular arm of the immune system; it can be transferred by CD4-positive T cells but not by serum. Infection with measles virus can suppress cell-mediated immunity, resulting in a loss of tuberculin skin test reactivity and, in some instances, reactivation of dormant organisms and clinical disease.
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A gene called Nramp determines natural resistance to tuberculosis. People who have mutations in the Nramp gene have a much higher rate of clinical tuberculosis than those with a normal allele. The NRAMP protein is located in the membrane of the phagosome in macrophages and plays an important role in killing the organism within the phagosome.
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The clinical findings are varied, and many organs can be involved, but the lungs are the main site of infection. Constitutional symptoms such as fever, fatigue, night sweats, and weight loss are common.
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In pulmonary tuberculosis, the main findings are cough and hemoptysis. The chest X-ray findings in reactivation tuberculosis of the lung include an infiltrate in the upper lobe with or without a cavity.
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Scrofula is mycobacterial cervical lymphadenitis that presents as swollen, nontender lymph nodes, usually unilaterally. M. tuberculosis causes most cases of scrofula, but nontuberculous mycobacteria (NTM), such as Mycobacterium scrofulaceum, can also cause scrofula. Lymphadenitis is the most common extrapulmonary manifestation of tuberculosis. Patients infected with human immunodeficiency virus (HIV) are more likely to have multifocal lymphadenitis than those not infected with HIV.
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Erythema nodosum, characterized by tender nodules along the extensor surfaces of the tibia and ulna, is a manifestation of primary infection seen in patients who are controlling the infection with a potent cell-mediated response (Figure 21–4). Miliary tuberculosis is characterized by multiple disseminated lesions that resemble millet seeds. Tuberculous meningitis and tuberculous osteomyelitis, especially vertebral osteomyelitis (Pott’s disease), are important disseminated forms.
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Gastrointestinal tuberculosis is characterized by abdominal pain and diarrhea accompanied by more generalized symptoms of fever and weight loss. Intestinal obstruction or hemorrhage may occur. The ileocecal region is the site most often involved. Tuberculosis of the gastrointestinal tract can be caused by either M. tuberculosis when it is swallowed after being coughed up from a lung lesion or by M. bovis when it is ingested in unpasteurized milk products. Oropharyngeal tuberculosis typically presents as a painless ulcer accompanied by local adenopathy.
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In renal tuberculosis, dysuria, hematuria, and flank pain occur. “Sterile pyuria” is a characteristic finding. The urine contains white blood cells, but cultures for the common urinary tract bacterial pathogens show no growth. However, mycobacterial cultures are often positive.
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Note that most (approximately 90%) infections with M. tuberculosis are asymptomatic. Asymptomatic infections, also known as latent infections, can reactivate and cause symptomatic tuberculosis. The most important determinant of whether overt disease occurs is the adequacy of the host’s cell-mediated immune (CMI) response. For example, AIDS patients have a very high rate of reactivation of prior asymptomatic infection and of rapid progression of the disease. In these patients, untreated disease caused by M. tuberculosis has a 50% mortality rate.
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Furthermore, administration of infliximab (Remicade), a monoclonal antibody that neutralizes tumor necrosis factor (TNF), has activated latent tuberculosis in some patients. The explanation for this is that TNF activates helper T cells to make gamma-interferon that activates macrophages to kill M. tuberculosis. So if TNF is neutralized by infliximab then gamma-interferon is not made and killing of M. tuberculosis by macrophages is reduced. Remicade is used in the treatment of rheumatoid arthritis (see Chapter 66). Diabetics are also predisposed to reactivation and progression of disease.
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In some patients with AIDS who are infected with M. tuberculosis, treating the patient with highly active antiretroviral therapy (HAART) causes an exacerbation of symptoms. This phenomenon is called immune reconstitution inflammatory syndrome (IRIS). The explanation of the exacerbation of symptoms is that HAART increases the number of CD4 cells, which increases the inflammatory response. To prevent this, patients should be treated for the underlying infection before starting HAART.
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Asymptomatic adults who are at high risk of having a latent infection should be screened using either the PPD skin test or the interferon-γ release assay (IGRA) test. Examples of adults at high risk are those who have lived in countries with increased prevalence of tuberculosis and those who are homeless. If found to be positive, these patients should be treated for latent infection. Screening tests and the treatment of latent infections are described later in this chapter.
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Acid-fast staining of sputum or other specimens is the usual initial test (see Figure 21–1). Either the Kinyoun version of the acid-fast stain or the older Ziehl-Neelsen version can be used. The acid-fast stain has low sensitivity, as evidenced by the finding that approximately 50% of “smear-negative” samples are “culture-positive.” For rapid screening purposes, auramine stain, which can be visualized by fluorescence microscopy, is used.
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In addition to performing an acid-fast stain, the specimen should be cultured. After digestion of the specimen by treatment with NaOH and concentration by centrifugation, the material is cultured on special media, such as Löwenstein-Jensen agar or Middlebrook agar, for up to 8 weeks. It will not grow on a blood agar plate. In liquid BACTEC medium, radioactive metabolites are present, and growth can be detected by the production of radioactive carbon dioxide in about 2 weeks. A liquid medium is preferred for isolation because the organism grows more rapidly and reliably than it does on agar. If growth in the culture occurs, the organism can be identified by biochemical tests. For example, M. tuberculosis produces niacin, whereas almost no other mycobacteria do. It also produces catalase.
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Nucleic acid amplification tests (NAATs) can be used to detect the presence of M. tuberculosis directly in clinical specimens such as sputum. NAATs are available that detect either the ribosomal RNA or the DNA of the organism. These tests are highly specific, but their sensitivity varies. In sputum specimens that are acid-fast stain positive, the sensitivity is high, but in “smear-negative” sputums, the sensitivity is significantly lower. These tests are quite useful in deciding whether to initiate therapy prior to obtaining the culture results.
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Because drug resistance, especially to isoniazid (see later), is a problem, susceptibility tests should be performed. However, the organism grows very slowly, and susceptibility tests usually take several weeks, which is too long to guide the initial choice of drugs. To address this problem, molecular tests are available that detect mutations in the chromosomal genes that encode either the catalase gene, which mediates resistance to isoniazid, or the RNA polymerase gene, which mediates resistance to rifampin. Whole genome sequencing can detect genotypic changes that determine resistance to isoniazid, rifampin, ethambutol, and pyrazinamide.
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A urine test for active tuberculosis is available. The test detects the lipoarabinomannan antigen with high sensitivity and specificity. Its main drawback is that it does not provide drug susceptibility, so additional testing is required.
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The luciferase assay, which can detect drug-resistant organisms in a few days, is also used. Luciferase is an enzyme isolated from fireflies that produces flashes of light in the presence of adenosine triphosphate (ATP). If the organism isolated from the patient is resistant, it will not be damaged by the drug (i.e., it will make a normal amount of ATP), and the luciferase will produce the normal amount of light. If the organism is sensitive to the drug, less ATP will be made and less light produced.
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There are two approaches to the diagnosis of latent infections. One is the PPD skin test as described in the “Immunity & Hypersensitivity” section earlier in this chapter. Because there are problems both in the interpretation of the PPD test and with the person returning for the skin test to be read, a quantifiable laboratory-based test is valuable.
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This laboratory test is an interferon-γ release assay (IGRA), and there are two versions available: QuantiFERON-TB Gold and T-SPOT.TB. In the IGRA assay, blood cells from the patient are exposed to antigens from M. tuberculosis, and the amount of interferon-γ released from the cells is measured. The sensitivity and specificity of the IGRA are as good as those of the PPD skin test. Because the antigens used in the test are specific for M. tuberculosis and are not present in BCG, the test is not influenced by whether a person has been previously immunized with the BCG vaccine.
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Note that the IGRA and PPD tests are positive in both latent disease and in active tuberculosis, so any person with a positive test must be evaluated for the presence of active disease by obtaining a chest X-ray and a sputum sample.
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Treatment & Resistance
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Multidrug therapy is used to prevent the emergence of drug-resistant mutants during the long (6- to 9-month) duration of treatment. (Organisms that become resistant to one drug will be inhibited by the other.) Isoniazid (INH), a bactericidal drug, is the mainstay of treatment. Treatment for most patients with pulmonary tuberculosis is with three drugs: INH, rifampin, and pyrazinamide. INH and rifampin are given for 6 months, but pyrazinamide treatment is stopped after 2 months. A somewhat different regimen can also be used. A convenient way to remember that regimen is to give four drugs (isoniazid, rifampin, pyrazinamide, and ethambutol) for 2 months and two drugs (isoniazid and rifampin) for 4 months. In patients who are immunocompromised (e.g., AIDS patients), who have disseminated disease, or who are likely to have INH-resistant organisms, a fourth drug, ethambutol, is added, and all four drugs are given for 9 to 12 months.
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Although therapy is usually given for months, the patient’s sputum becomes noninfectious within 2 to 3 weeks. The necessity for protracted therapy is attributed to (1) the intracellular location of the organism; (2) caseous material, which blocks penetration by the drug; (3) the slow growth of the organism; and (4) metabolically inactive “persisters” within the lesion. Because metabolically inactive organisms may not be killed by antitubercular drugs, treatment may not eradicate the infection, and reactivation of the disease may occur in the future.
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Lymphadenitis, including cervical lymphadenitis (scrofula) caused by M. tuberculosis, should be treated with the drug regimens described earlier for disseminated disease. Scrofula caused by M. scrofulaceum can be treated by surgical excision of the single cervical lymph node, but alternative approaches exist. A complete discussion of these is beyond the scope of this book.
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Treatment of latent (asymptomatic) infections consists of INH taken for 6 to 9 months or INH plus rifapentine for 3 months. A course of rifampin for 4 months is also effective. This approach is most often used in asymptomatic patients whose PPD skin test or IGRA test recently converted to positive. The risk of symptomatic infection is greatest within the first 2 years after infection, so INH is particularly indicated for these “recent converters.” INH is also used in children exposed to patients with symptomatic tuberculosis. Patients who receive INH should be evaluated for drug-induced hepatitis, especially those over the age of 35 years. Rifampin can be used in those exposed to INH-resistant strains. A combination of rifampin and pyrazinamide should not be used because it causes a high rate of severe liver injury.
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Resistance to INH and other antituberculosis drugs is being seen with increasing frequency in the United States, especially in immigrants from Southeast Asia and Latin America. Strains of M. tuberculosis resistant to multiple drugs (MDR strains) have emerged, primarily in AIDS patients. The most common pattern is resistance to both INH and rifampin, but some isolates are resistant to three or more drugs. The treatment of MDR organisms usually involves the use of at least four drugs, including ciprofloxacin, amikacin, ethionamide, and cycloserine. In 2019, the FDA approved a three drug combination for the treatment of MDR tuberculosis consisting of pretomanid, linezolid, and bedaquiline. The precise recommendations depend on the resistance pattern of the isolate and are beyond the scope of this book.
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In 2013, bedaquiline was approved for the treatment of MDR strains. It should be used in combination with other drugs, not as monotherapy. It is a diarylquinoline that inhibits an ATP synthase unique to M. tuberculosis.
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Previous treatment for tuberculosis predisposes to the selection of these MDR organisms. Noncompliance (i.e., the failure of patients to complete the full course of therapy) is a major factor in allowing the resistant organisms to survive. One approach to the problem of noncompliance is directly observed therapy (DOT), in which health-care workers observe the patient taking the medication.
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The strains of M. tuberculosis resistant to INH, rifampin, a fluoroquinolone, and at least one additional drug are called extensively drug-resistant (XDR) strains. XDR strains emerged in 2005 among HIV-infected patients in South Africa.
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Note that M. tuberculosis produces β-lactamase, rendering the organism resistant to many penicillins and cephalosporins. Trials using amoxicillin-clavulanate to treat active tuberculosis were unsuccessful.
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The incidence of tuberculosis began to decrease markedly even before the advent of drug therapy in the 1940s. This is attributed to better housing and nutrition, which have improved host resistance. At present, prevention of the spread of the organism depends largely on the prompt identification and adequate treatment of patients who are coughing up the organism. The use of masks and other respiratory isolation procedures to prevent spread to medical personnel is also important. Contact tracing of individuals exposed to patients with active pulmonary disease who are coughing should be done.
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An important component of prevention is the use of the PPD skin test to detect recent converters and to institute treatment for latent infections as described earlier. Groups that should be screened with the PPD skin test include people with HIV infection, close contacts of patients with active tuberculosis, low-income populations, alcoholics and intravenous drug users, prison inmates, and foreign-born individuals from countries with a high incidence of tuberculosis.
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Because there are some problems associated with PPD skin tests, such as the measurement and the interpretation of results and the inconvenience of the patient having to return for the skin test to be read, a laboratory test to detect latent infections was developed. This test, called QuantiFERON-TB Gold (QFT-G), measures the amount of interferon-γ released from the patient’s lymphocytes after exposure to antigens from M. tuberculosis in cell culture. QFT-G requires only a single blood specimen and determines the amount of interferon-γ by an enzyme-linked immunosorbent assay (ELISA) test.
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BCG vaccine can be used to induce partial resistance to tuberculosis. The vaccine contains a strain of live, attenuated M. bovis called bacillus Calmette-Guérin. The vaccine is effective in preventing the appearance of tuberculosis as a clinical disease, especially in children, although it does not prevent infection by M. tuberculosis. However, a major problem with the vaccine is its variable effectiveness, which can range from 0% to 70%. It is used primarily in areas of the world where the incidence of the disease is high. It is not usually used in the United States because of its variable effectiveness and because the incidence of the disease is low enough that it is not cost-effective.
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The skin test reactivity induced by the vaccine given to children wanes with time, and the interpretation of the skin test reaction in adults is not altered by the vaccine. For example, skin test reactions of 10 mm or more should not be attributed to the vaccine unless it was administered recently. In the United States, use of the vaccine is limited to young children who are in close contact with individuals with active tuberculosis and to military personnel. BCG vaccine should not be given to immunocompromised people because the live BCG organisms can cause disseminated disease.
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BCG vaccine is also used to treat bladder cancer. The vaccine is instilled into the bladder and serves to nonspecifically stimulate cell-mediated immunity, which can inhibit the growth of the carcinoma cells.
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As an alternative to BCG vaccine, a trial vaccine containing two recombinant M. tuberculosis proteins as immunogen was shown to be effective in preventing active disease.
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Pasteurization of milk and destruction of infected cattle are important in preventing intestinal tuberculosis.