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    Home > Active Ingredient News > Infection > Rapid identification of two cases of AIDS-combined fungal blood flow infection analysis

    Rapid identification of two cases of AIDS-combined fungal blood flow infection analysis

    • Last Update: 2020-06-24
    • Source: Internet
    • Author: User
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    Acquiredimmunodeficiencysyndrome (AIDS) is an infectious disease caused by humanimmunodeficiency virus (HIV), which is characterized by damage to the immune system, and patients are highly susceptible toopportunisticinfection, while opportunistic infections of deep fungi such as new cryptococcal bacteria and Malnifi blue bacteria are one of the leading causes of death in AIDS patientsThe most common lysacious infection caused by new types of cryptococcal bacteria is cryptococcal meningitis, which has a hidden incidence and a high fatality ratePatients are mainly characterized by unbearable headaches, accompanied by fever, nausea, vomiting, meninges stimulation positive and other symptoms and signs of meningitisCryptococcal meningitis is rare in healthy populations, but is more common in patients with AIDSMalnifi blue bacteria is a highly pathogenic fungus, but also the only two-way fungus in the genico of penicillin, mainly affecting immunocompromised populations, especially AIDS patientsIts clinical manifestations are complex and diverse and lack specific, and it has been reported that 85% of Malnifi blue bacteria infections occur in patients with AIDS and have become one of the indicative symptoms of THE diagnosis of AIDS clinicalThe matrix-assisted laser resolution ionization flight time mass spectrometry (MALDI TOF MS) is a new soft ionizing biomass spectrum technology, which detects the protein fingerprint map of the characteristics of each pathogen and compares it with the map library in the database, i.eit can identify the pathogen to be testedThe direct detection of pathogens in blood-positive bottles by combining MALDI-TOF MS gelding tubes can greatly shorten the detection time and provide a reliable clinical basis for diagnosisThis paper reviews and analyzes 2 clearly diagnosed cases of combined fungal infection of AIDS, and explores the clinical value of mass spectrometry identification for the diagnosis of combined bacterial infection in AIDS patients1 case data1General information In September 2018, 2 cases of HIV patients infected with combined fungal infections were admitted to the respiratory and haematology departments of changzhou First People's HospitalCase 1, male, 56 years old, due to "cough with weakness, more than half a month of difference" was admitted to hospitalPatients admitted to hospital in January without an incentive to appear a burst of cough, 1 week ago to the local hospital to check the chest tablets promptbronitis, to "Shunning, Su Huang cough capsules", the symptoms did not improve, after the emergence of fatigue, nacho sexual aggravation2 d before eating after vomiting a little stomach contents, in this hospital consultation in parallel related examinationThe patient has gradually reduced his or her self-reported self-in-the-disease and reduced his body mass by about 5 kgThe patient is clear-minded, mentally withered, chronically illChest CT shows the right lung lower lobe multiple cavity lesions, multiple lung large herpes with infection, the right lung lower leaf outer substrate section nodules, the upper lobe of the right lung after a little inflammation, left lung multi-fiber stoveDuring the course of the patient appears a seizure of the eyes, lasting a few seconds after the improvement; Diagnosed as: lung shadow: fungal infection possible; encephalitis (fungal possible)Case 2, female, 39, was admitted to hospital after "discovering a swollen lymph node on the collarbone for more than 2 weeks"Patients 2 weeks ago no obvious cause of the left clavicle on the lymph nodes sore swollen, accompanied by coughing, coughing sputum, fever, and feeling weak, dizziness and discomfortThe patient had fainted 1 time during the course of the disease and regained consciousness after about 15 sThe patient's body temperature is 39 degrees C, the mind is clear, the spirit is wiltingThe positive electron emission computer tomography showed an increase in the metabolism of multiple lymphunofluororide deoxyglucose, consideringlymphoma immersion; The diagnosis is: lymph node swelling to be checked, lung infection 1 2 MALDI -TOF MS Identification Extract 5 mL positive blood culture fluid injected into the isolated gel coagulation tube, oxygen bottle extracted blood culture with 4 000 r / mi n centrifugal 10 mi n, anaerobic bottle extracted culture liquid by 3 500 r / mi n centrifugal 7 mi n, separation of the pathogen to be measured Discard the separation of the upper liquid inside the gel-promoting tube, and slowly inject 1 mL of sterile distilled water into the sediment already suspended (does not destroy the gel), transferring the suspension to the bacteria-free 1 5 mL small centrifuge tube, by 13 000 r /mi n centrifugal 1 mi n Discard the upper liquid and repeat the washing once, and finally the bacteria at the bottom of the tube after centrifugation is suspended in 1 mL sterile distilled water for inspection Take 300 sl of the bacteria resuspended with 900 sl aqueous ethanol, discard the upper liquid after 13000 r/mi n centrifugal 1 mi n, and then after multiple centrifugation to completely remove the remaining waterless ethanol, placed in the air naturally dry Add 70% totherin acid 30 sL to fully mix the bacteria to be tested, then add acetylene 30 sL vortex mix, after 13 000 r /mi n centrifugal 2 mi n, take the upper liquid for MALDI -TOF MS identification Take the prepared upper clear liquid 1 sl coating on the 96-well metal target plate, after room temperature drying in the metal target plate surface added matrix liquid alpha - cyanide 4 - hydroxyl cinnamon acid 1 sL, again after room temperature drying for testing Parameter settings: linear, positive ions, protein peak spectrum m/z range 2 to 20 x 10 3, laser resolution 100 times per hole App B i o t ype r 3 0 all progress number according to the library comparison, to determine the result 1 3 API 20 C slat identification Extraction of culture liquid in a blood culture bottle for pure culture, the isolated colonies formulated with sterile physiological saline into 2 McKeaus units to be tested, take 100 ?L suspension to the API CMed i um medium, mixed and added the identification plate, 28 degrees C culture 72 h for interpretation of the results 1 4 Fungal small culture The potato glucose agar medium is cut into small pieces placed on the slide, with the vaccination ring to pick pure culture of silky bacteria inoculated in the four corners of the medium, covered with a glass, 28 degrees C culture 2 to 3 d after dyeing with lactic acid mamphione, under the hyperscope to observe the fungal form Note: A indicates the results of the direct smear of the Gramn staining mirror (10 x 100); B indicates the incubating 48 h colony form at 35 degrees C; C indicates the incubating 48 h colony form at 28 degrees C; D represents fungal small culture lactic acid medathol staining mirror results (10 x 40) Figure 3 Case 2 Blood Culture Positive Smear Results and Pure Culture Colony Characteristics and Mirror Results 2 Results 2.1 Mass Spectrometry Identification Results 2 patients were sent to blood training after admission, and were directly tested with mass spectrometry after blood culture was reported to yang Case 1 identification results were for new erythroccid bacteria with a mass spectrometry score of 2 142。 Case 2 identification results were found to be some species of penicillin, with a mass spectrometry score of 1 474。 See Figure 1 2 2 Traditional identification results Case 1: The patient's blood culture two-sided oxygen bottle was positive at 67, 70 h, respectively, the smear gramgelan dye mirror test found positive round spores The specimens were inoculated on a blood agar plate, chocolate agar plate, and sand-protected weak medium, and the cultures of 35 degrees C 48 h all grew white smooth yeast-like colonies The API 20 C slats were identified as new cryptococcal bacteria Figure 2 Case 2: The patient's blood culture two-sided oxygen bottle sydd at 77, 89 h alarm positive, smear editing gel found rich silk and a small amount of fungal spores, spores two blunt circle, the middle of the separated sausage shape The specimens were inoculated on the blood agar plate, chocolate agar plate and sand weak medium, and incubated 48 h at 28 degrees C and 35 degrees C respectively, and the gray-white wax samples, membrane-like flat colonies were visible on the blood flat plate, and the center of the colony was visible in the umbilical depression; White yeast-like colonies can be seen on the weak slope dede of the sand, which is cultured at 35 degrees C, and does not produce red pigment The light red fluffy colonies can be seen on the sand-preserved weak slope surface cultured at 28 degrees C, and the entire medium is dyed to rose red Fungal small culture can be seen branch-separated mycelium, spores stalks light slip, is a crucified branch dispersion, mostly symmetrical double-wheeled; The biphase and crucible-shaped branches of culture can be identified as Marnifi blue bacteria Figures 2, 3 2 3 Final diagnosis Case 1 Patients after consultation with neurology after considering brain vascular accident and HIV encephalopathy, after consultation with the city infectious diseasehospital to consider AIDS, central nervous system infection, cryptococcal encephalitis To glycol dehydration, Vulcan static drop anti-fungal and correct electrolyte disorders and other treatment, and transferred to the city infectious diseases hospital for further treatment, the current patient is stable and recovering well Case 2 patients lymph node biopsy showed the growth of lymphatic tissue on the collarbone, neutrophil sciatica immersion, small abscess estoms accompanied by a multi-volume tissue cell reaction, tending to infection changes Ask the city infectious disease hospital to consider AIDS, and transfer to the city infectious diseases hospital for further treatment Note: A indicates the results of the direct smear of the Gramn staining mirror (10 x 100); B indicates the incubating 48 h colony form at 35 degrees C; C indicates the incubating 48 h colony form at 28 degrees C; D represents fungal small culture lactic acid medathol staining mirror results (10 x 40) Figure 3 Case 2 Blood Culture Positive Smear Results and Pure Culture Colony Characteristics and Mirror Results 3 Discussion Fungal Blood Flow Infection is a more common opportunistic infection in AIDS patients, Xie Chaoyun and others reported the incidence of AIDS cocombinal fungal infection s36.61% The new erythrococcal bacteria and the Malnifi blue bacteria are commonly used in AIDS disease-causing fungi, and there are literature reports of a one-year attributable mortality rate of 24 2% of the disease death rate of co-infection with Malnifi blue bacteria can reach 18% to 26%, so timely etiology diagnosis is the key Cryptococcal meningitis patients are more symptoms of the central nervous system such as fever, headache and vomiting, case 1 patients in the course of the disease, neck slight resistance and other cryptococcal meningitis-related symptoms, and the final identification results are fully consistent After Malnifi blue bacteria infection, clinical manifestations of fever, large liver and spleen, lymph node swelling, breathing and digestive system abnormal symptoms, patients in case 2 by lymph node biopsy prompted lymph node abscesses, excluding the possibility of lymphoma, its symptoms are consistent with Malnifi blue bacteria infection Review the diagnosis process of 2 patients, patients in case 1 suspected infection, case 2 patients are more complex, high fever and the main complaint of gonorrhea swollen, for these two types of clinically temporarily undiagnosed patients, first of all, blood, cerebrospinal fluid, bone marrow and other specimens should be collected for smearing and training, as far as possible to find the pathogen basis, timely pathogen diagnosis for the successful treatment of deep fungal infectionplay play a great role Although the traditional method of fungal identification is more accurate, it takes a long time and requires high experience on the tester, which can easily lead to misdiagnosis and misdiagnosis The use of mass spectrometers greatly shortens the detection time, the separation of gel-promoting coagulation joint mass spectrometer can directly detect blood culture-positive pathogens, shortenthe time for transspecies culture identification The traditional identification of cryptococcal bacteria mainly depends on API slats, it takes 72 h to interpret the results, and ink dyeing is more convenient, but because of the thickness and thinness of the cryptococcal pod, the results of the observation of the experience of the subject is greater The traditional method of identifying the Malnifi blue bacteria requires 4 to 10 d , using the method of rapid identification of mass spectrometry can give the preliminary identification results of the genus on the same day, and the test results are consistent with the results of traditional fungal culture, greatly reducing the time spent on the cogal identification cost Although the level of penicillin can only be identified to the genus, still need to combine classic fungal culture and morphological observation to identify the level of species, but this can undoubtedly for the clinical diagnosis and treatment to gain more time In addition, compared with traditional fungal identification methods, the use of mass spectrometers to directly identify the blood culture positive bottle of fungal operation is simpler, and easy to develop standardized operating procedures, thereby reducing the identification error caused by human factors In summary, AIDS patients are prone to combined conditions of pathogenic fungal infection, the diagnosis of fungal infection depends on pathogen identification Traditional morphological identification is important for the diagnosis of fungal infection, but it takes a long time, and the emergence of mass spectrometry technology simplifies the operation process, shortens the identification time, and wins the time for the treatment of clinical fungal infection Improve the blood culture and delivery rate of patients with unknown causes of fever or infection, find pathogens as early as possible and treat the disease in a timely manner, combine with emerging mass spectrometry technology to shorten the detection time of pathogens, and provide a more timely and reliable basis for clinical diagnosis references
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