Cryptococcal Antigen Research Articles

Evaluation of the Biofire FilmArray meningitis/encephalitis assay for the detection of Cryptococcus neoformans/gattii

ObjectivesCryptococcal meningitis (CM) remains an important cause of morbidity and mortality among immunocompromised patients. Laboratory diagnostics for CM includes antigen detection, staining and culture. Data on the performance of the Biofire® FilmArray® meningitis/encephalitis (ME) panel for detecting Cryptococcus neoformans/gattii is limited, with several reports describing false negativity for this target. MethodsA retrospective analysis of 1384 physician-ordered ME panel tests ordered between January 2017 to October 2018 was performed. ME panel results were compared to cerebrospinal fluid (CSF) cryptococcal antigen (CrAg) and CSF culture testing and clinical significance of cryptococcal detection was determined. ResultsThere were 34 patients positive for cryptococcal detection by either ME panel, CSF CrAg or CSF culture in 2.7% of CSF specimens tested (38/1384). Of the 34 patients positive for cryptococcal detection, 85.3% were human immunodeficiency virus positive (29/34). The ME panel detected 32/38 (84.2%) cryptococcal-positive specimens, culture detected 28/38 (73.7%) and CSF CrAg was positive in 37/38 specimens (97.4%). The ME panel had a sensitivity and specificity of 96.4% (95% CI 81.7–99.9%) and 99.6% (95% CI 99.2–99.9%) compared with culture, and 83.8% (95% CI 68.0–93.8%) and 99.9% (95% CI 99.6–100.0%) compared to CSF CrAg testing, respectively. CrAg titres were lower among ME panel-negative, culture-negative specimens compared with ME panel-positive, culture-negative specimens (reciprocal median end-point titres of 128 ± 60 vs. 1920 ± 1730, p 0.04). All five CrAg-positive, ME panel- and culture-negative specimens were obtained from previously treated CM patients. DiscussionThe ME panel had high correlation with CSF culture and a somewhat lower correlation with CSF CrAg testing. The potential utility of using negative ME panel test results to predict culture sterility among patients undergoing treatment for CM warrants further study.

Epidemiological characteristics of cryptococcal meningoencephalitis associated with Cryptococcus neoformans var. grubii from HIV-infected patients in Madagascar: A cross-sectional study.

Cryptococcal meningoencephalitis (CM) remains the most prevalent invasive fungal infection worldwide. The main objective of this study was to describe the prevalence of CM and cryptococcal infection in HIV-infected patients in Madagascar. The secondary objectives were to assess the adjusted prevalence of CM according to clinical presentation and patient characteristics, to determine crude 90-day survival according to cryptococcal antigen (CrAg) status and CM, and to identify the genotypes of Cryptococcus clinical isolates. This cross-sectional study was carried out at two urban hospitals in Antananarivo (central highlands) and Toamasina (east coast) between November 2014 and December 2016. Consecutive HIV-infected adults presenting with CD4 cell counts ≤200/μl were enrolled. Lateral flow immunoassays of CrAg were performed on serum for all patients, and on cerebrospinal fluid for patients with CM symptoms. MALDI-ToF MS, ITS sequencing, and determinations of the molecular and mating types of the isolates were performed. Fluconazole is the only drug for CM treatment available in Madagascar. Patients were treated orally, with high doses (1200 mg/day) for 10–12 weeks and then with 200 mg/day. Minimum inhibitory concentrations were determined for amphotericin B, flucytosine, voriconazole and fluconazole in E-tests. Overall prevalence was 13.2% (95% CI 7.9–20.3) for cryptococcal infection and 10.9% (95% CI 6.1–17.5) for CM, among the 129 HIV-infected patients studied. The 90-day mortality rate was 58.8% (10/17) in CrAg-positive patients and 17.9% (20/112) in CrAg-negative patients (p<0.001). The 13 Cryptococcus strains obtained at baseline were all Cryptococcus neoformans var. grubii, genotypes VNI-αA (3 isolates), VNII-αA (4 isolates) or hybrid VNI/VNII-αAAα (6 isolates), suggesting high diversity. Two strains acquired fluconazole resistance after four and five months of exposure, respectively. The prevalence of cryptococcosis is high in Madagascar and this serious condition is life-threatening in HIV-infected patients. These findings will be used to raise the awareness of national authorities to strengthen the national HIV/AIDS control program.

Prevalence of cryptococcal meningitis in a tertiary care centre.

Background: Cryptococcal meningitis has emerged as an important opportunistic central nervous system (CNS) infection in Human Immunodeficiency Virus (HIV) positive patients. It is associated with a high mortality rate. Hence early diagnosis is necessary to start appropriate treatment. Cryptococcosis is generally found in association with acquired immunodeficiency syndrome (AIDS) although it has been reported to cause disease in HIV-seronegative patients also. Objective:Prevalence of Cryptococcal meningitis in a tertiary care centre. Material &amp; Methods: A total of 93 Cerebrospinal fluid samples from suspected cases of fungal meningitis were received in the microbiology department of GB Pant Hospital(GIPMER) from January to June 2018. Samples were subjected to direct microscopy- wet mount, India ink preparation and Gram stain, Cryptococcal antigen detection(Latex agglutination), and Fungal culture. In vitro susceptibility of Cryptococcus isolates to Fluconazole, Voriconazole, 5- Flucytosine and Amphotericin B was performed using standard broth microdilution method. Results: Out of 93 CSF samples, 6 were positive for India ink preparation and showed gram positive budding yeast cells by gram staining. All 6 samples were positive for the cryptococcal antigen test and fungal culture. Five were identified as C. neoformans and one C. gatti. All strains were susceptible to Amphotericin B. Four patients were HIV reactive and succumbed to the diseaseduring treatment. Two patients were found to be coinfected with Hepatitis B virus. Ashish Bajaj1, Bibhabati Mishra2, Poonam S Loomba2, Archana Thakur3, Abha Sharma4, Prachala G Rathod1, Madhusmita Das1, Ashna Bhasin1 1Senior Resident, 2Director Professor, 3Director Professor &amp; Head, 4Assistant Professor, Department of Microbiology, Govind Ballabh Pant Institute of Postgraduate Medical Education and Research (GIPMER), New Delhi, India ABSTRACT Cryptococcal meningitis has emerged as an important opportunistic central nervous system (CNS) infection in immunosuppressed patients.It is associated with a high mortality rate(&gt;30%) in immunosuppressed patients especially those infected with HIV, hence early diagnosis is necessary to start appropriate treatment. Prevalence of cryptococcal infection is increasing in developing nations including India as per some recent studies.(1) Conclusion: Index of suspicion of Cryptococcus infection as a possible cause of meningitis must be considered in chronic meningitis cases. Microscopy (India ink preparation) may be used as a cheap and rapid diagnostic tool.

Cryptococcus neoformans meningitis in an immunocompetent adult: A case report

Neuromeningeal cryptococcosis (NMC) is relatively rare in immunocompetent subjects. We report a Congolese case of NMC diagnosed in an apparently immunocompetent subject. A 35-year-old woman was admitted for acute meningoencephalitis. The cerebrospinal fluid (CSF) examination revealed cytological and biochemical abnormalities and the presence of Cryptococcus neoformans on direct examination with India ink and in culture. The serum dosage of cryptococcal antigens was titrated at 1:32. HIV and syphilis antibodies were negative and the blood CD4 lymphocyte count was 610/mm3. The patient had no other immunosuppressive factors, in particular no hematologic abnormalities, no solid tumor, or malnutrition. She was unsuccessfully treated with fluconazole intraveinously, and died after 10 days of treatment. This observation reminds us that NMC can occur in subjects without apparent immunosuppressive factor with a misleading clinical presentation. It is a rare infection with a severe prognosis even in immunocompetent patients.

Should cryptococcal antigen screening be considered as a routine procedure in antiretroviral therapy naïve severely immunocompromised HIV-seropositives – a prevalence study from Eastern India to support recent 2018 WHO guidelines

Should cryptococcal antigen screening be considered as a routine procedure in antiretroviral therapy naïve severely immunocompromised HIV-seropositives – a prevalence study from Eastern India to support recent 2018 WHO guidelines

Disseminated and cutaneous cryptococcosis by C. neoformans VNI in an immunocompetent patient

ABSTRACT Cryptococcosis is caused by yeasts of the Cryptococcus neoformans/C. gattii complex, presenting cutaneous, respiratory and disseminated forms. A 44-year-old immunocompetent male with facial lesion and latent pneumonia was hospitalized and misdiagnosed with paracoccidioidomycosis. Computerized tomography scans showed pulmonary and neurological involvement, and cultures/China ink, cryptococcal antigen test and restriction fragment length polymorphism of urease gene (URA5-RFLP) confirmed C. neoformans genotype VNI. Hemoculture indicated ampicillin-resistant Klebsiella pneumoniae (healthcare-associated infection). Fluconazole was administered, but after resistance detection, amphotericin B was chosen (cumulative dose/1500 mg). The patient was discharged with clinical remission (75 days) and amphotericin for one year (maintenance phase).

Cryptic pulmonary cryptococcosis: An atypical presentation of pulmonary cryptococcosis in a HIV-infected patient

Pulmonary cryptococcosis is a fungal infection caused by inhalation of Cryptococcus gattii and/or C. neoformans spores. It mostly affects HIV-infected patients. This is a case report of a severely immunocompromised HIV-infected patient, presenting with respiratory symptoms and atypical chest X-ray features for pulmonary cryptococcosis. A serum cryptococcal latex antigen test is positive in a majority of HIV-infected patients with pulmonary cryptococcosis. This case report demonstrates the occurrence of a false negative serum cryptococcal latex antigen test, becoming positive with the development of an unmasking immune reconstitution syndrome (after antiretroviral therapy was commenced). This also resulted in the characteristic cryptococcal lung cavities observed on computed tomography chest images. Duration of fluconazole therapy should be individualised, and serial chest imaging (e.g. chest X-ray) should be performed to monitor treatment response.

An Unusual Cause of Recurrent Pleural Effusion

Abstract Cryptococcus neoformans is a yeast that can cause severe infection in the immunocompromised. It is commonly associated with meningitis in patients with underlying HIV infection; however, infections can occur in other immunosuppressed hosts as well as the immunocompetent. Pulmonary disease is well described, but recurrent pleural effusion and empyema are rare. We describe a case in a non–HIV-infected, nontransplant cirrhotic patient presenting with recurrent pleural effusions and cryptococcal empyema. The patient is a 66-year-old man with cirrhosis, diabetes, and kidney disease who presented with 3 months of dyspnea complicated by recurrent pleural effusion and low protein ascites after treatment of hepatitis C virus. Routine cultures from multiple thoracenteses were unrevealing. Fungal culture ultimately grew C. neoformans/gattii. His serum cryptococcal antigen titer was 1:40, and chest imaging revealed several pulmonary nodules. He was admitted for evaluation of disseminated disease and central nervous system involvement, which was unremarkable. He was started on oral fluconazole 200 mg daily dosed for his renal function and was doing well in outpatient follow-up. Patients with cirrhosis tend to have a higher incidence of cryptococcal disease and are more likely to have disseminated infection with higher mortality. It is possible that deficiencies in complement and cytokine levels as well as leukocyte function may contribute to this increased risk. Effusions are an uncommon manifestation of pulmonary cryptococcosis and, as with the case in our patient, may resemble hepatic hydrothorax, leading to a delay in antifungal therapy. In conclusion, patients with liver disease have an increased risk of cryptococcosis. A high index of suspicion must be maintained for infection and should be considered in patients with recurrent effusions.

Clinical diversity of invasive cryptococcosis in AIDS patients from central China: report of two cases with review of literature

BackgroundAlthough antiretroviral therapy (ART) has greatly improved the prognosis of acquired immunodeficiency syndrome (AIDS) patients globally, opportunistic infections (OIs) are still common in Chinese AIDS patients, especially cryptococcosis.Case presentationWe described here two Chinese AIDS patients with cryptococcal infections. Case one was a fifty-year-old male. At admission, he was conscious and oriented, with papulonodular and umbilicated skin lesions, some with ulceration and central necrosis resembling molluscum contagiosum. The overall impression reminded us of talaromycosis: we therefore initiated empirical treatment with amphotericin B, even though the case history of this patient did not support such a diagnosis. On the second day of infusion, the patient complained of intermittent headache, but the brain CT revealed no abnormalities. On the third day, a lumbar puncture was performed. The cerebral spinal fluid (CSF) was turbid, with slightly increased pressure. India ink staining was positive, but the cryptococcus antigen latex agglutination test (CrAgLAT: IMMY, USA) was negative. Two days later, the blood culture showed a growth of Cryptococcus neoformans, and the same result came from the skin culture. We added fluconazole to the patient’s treatment, but unfortunately, he died three days later.Case two was a sixty-four-year-old female patient with mild fever, productive cough, dyspnea upon movement, and swelling in both lower limbs. The patient was empirically put on cotrimoxazole per os and moxifloxacin by infusion. A bronchofibroscopy was conducted with a fungal culture, showing growth of Cryptococcus laurentii colonies. Amphotericin B was started thereafter but discontinued three days later in favor of fluconazole 400 mg/d due to worsening renal function. The patient became afebrile after 72 h of treatment with considerable improvement of other comorbidities and was finally discharged with continuing oral antifungal therapy.ConclusionsOur cases illustrate that cryptococcal disease is an important consideration when treating immunocompromised individuals such as AIDS patients. Life threatening meningitis or meningoencephalitis caused by C. neoformansmay still common in these populations and can vary greatly in clinical presentations, especially with regard to skin lesions. Pulmonary cryptococcosis caused by C. laurentii is rare, but should also be considered in certain contexts. Guidelines for its earlier diagnosis, treatment and prophylaxis are needed.

A clinical rule for the prediction of meningitis in HIV patients in the era of combination antiretroviral therapy.

The diagnosis of meningitis in HIV patients is challenging due to altered immune responses. Diagnostic scoring systems were recently proposed for use in research settings to help prompt and easy differential diagnosis. The objective of this study was to create a clinical prediction rule (CPR) for meningitis in HIV-infected patients and to address the enigma of differentiating bacterial (BM), TB (TBM) and cryptococcal (CCM) meningitis based on clinical features alone, which may be enhanced by easy-to-obtain laboratory testing. We retrospectively enrolled 352 HIV patients presenting with neurological manifestations suggesting meningitis over the last 18 y (2000-2018). Relevant clinical and laboratory information were retrieved from inpatient records. The features independently predicting meningitis or its different types in microbiologically proven meningitis cases were modelled by multivariate logistic regression to create a CPR in an exploratory data set. The performance of the meningitis diagnostic score was assessed and validated in a subset of retrospective data. AIDS clinical stage, injecting drug use, jaundice and cryptococcal antigen seropositivity were equally important as classic meningitic symptoms in predicting meningitis. Arthralgia and elevated cerebrospinal fluid Lactate dehydrogenase (LDH) were strong predictors of BM. Patients with cryptococcal antigenemia had 25 times the probability of having CCM, whereas neurological deficits were highly suggestive of TBM. The proposed CPRs have good diagnostic potential and would support decision-making in resource-poor settings.

Cryptococcal antigenemia prevalence and clinical data in HIV-infected patients from the reference centre at INI-FIOCRUZ, Rio de Janeiro, Southeast of Brazil.

Cryptococcal meningitis is a several disease common in late stage of HIV infection. Detection of cryptococcal antigen (CrAg) is an important for early diagnosis of this invasive mycosis. The pre-emptive treatment for isolated antigenemia prevents the onset of meningoencephalitis. Screening CrAg in patients with low CD4 count is cost-effective in countries with prevalence of antigenemia above 3%. However, in Brazil, the number of prevalence studies on cryptococcosis and HIV is insufficient. The objective of this study is to estimate the prevalence of CrAg and describe clinical characteristics from a cohort of patients followed at a reference center in Brazil. CrAg screening was performed in 89 inpatients with CD4 count ≤200 cells/mm3 or WHO stage III/IV from the National Institute of Infecttious Disease, Rio de Janeiro. Patients with isolated antigenemia received pre-emptive therapy with fluconazole and patients with meningoencephalitis were treated with Amphotericin B. Individuals were followed up for 12 months. Prevalence of serum CrAg was 11.23%, cryptococcal meningoencephalitis 6.74% and isolated antigenemia 4.81%. None of the patients with isolated antigenemia developed meningoencephalitis during the follow up. Signs and symptoms of meningoencephalitis were unspecific or absent. Our study suggests the need of CrAg screening in Brazil and highlights that lumbar puncture is mandatory in all individuals CrAg positive to exclude asymptomatic meningoencephalitis.

Clinical characteristics of 81 cases with acquired immunodeficiency syndrome complicated with cryptococcal meningitis

Objective To investigate the clinical features and prognosis factors of acquired immunodeficiency syndrome (AIDS) patients complicated with cryptococcal meningitis (CM). Methods Retrospective analyses were performed on clinical features, laboratory data, treatment status and related prognosis factors in 81 AIDS patients with CM admitted to the Department of Infectious Diseases, Beijing You′an Hospital, Capital Medical University from January 2010 to December 2017. The t test, rank sum test and χ2 test were employed to analyze the data. Results Of the 81 AIDS patients with CM, 71 cases were infected with human immunodeficiency virus (HIV) by sexual transmission (87.7%). The most common clinical symptoms were fever in 60 (74.1%), headache in 72 (88.9%), and nausea and vomiting in 56 (69.1%). Cerebrospinal fluid (CSF) examination results show that 60 cases (74.1%) had elevated opening pressure, the white blood cell count was 17.0 (6.0, 44.5)×106/L, monocyte count was 9.0 (3.0, 29.5)×106/L, the level of chloride was (117.26±5.61) mmol/L, of glucose was 2.89 (2.05, 3.41) mmol/L, of protein was 0.32 (0.21, 0.65) g/L, ink staining positive rate was 84.0% (68/81), fungal culture positive rate was 59.3% (48/81). The positive rate of serum cryptococcal antigen was 96.3% (78/81), and CSF cryptococcal antigen positive rate was 93.8% (76/81). The clinical efficacies were not significant different among different treatment regimens (χ2=1.479, P=0.533). After treatment, 60 patients survived and 21 died, with an overall mortality rate of 25.9%. Univariate analysis showed that consciousness disorder and CSF opening pressure were significantly higher in the death group than those in the survival group (χ2=22.365, t=0.317, respectively, both P<0.05), while serum albumin (Alb) and CD4+ T lymphocyte counts were significantly lower in the death group than those in the survival group (t=7.975, Z=-3.073, respectively, both P<0.05). Multivariate logistic regression analysis showed that consciousness disorder and Alb were independent factors influencing the clinical outcome of AIDS patients with CM. Consciousness disorder was related with poor outcome (odd ratio (OR)=26.704, P=0.011, 95% confidence interval (CI) 2.115-337.247), and higher Alb was related with good outcome (OR=0.671, P=0.005, 95%CI 0.507-0.888). The area under the receiver operating characteristic curve of serum Alb for predicting poor outcomes of AIDS patients with CM was 0.932 (95% CI 0.859-0.998, P<0.01). By using 31.7 g/L as cut-off value, the sensitivity was 95% and the specificity was 81% for predicting poor outcome. Conclusions AIDS complicated with CM has a high mortality rate, and its clinical features are lack of specificity. Consciousness disorder and Alb are independent prognosis factors. Key words: Acquired immunodeficiency syndrome; Cryptococcus neoformans; Meningitis, cryptococcal; Clinical characteristics

Cerebrospinal fluid pleocytosis in immunocompromised patients: Can it be Cryptococcus.

Cryptococcal meningitis commonly affects immunocompromised cases and can have varied presentation. In some instances, the presence of a plethora of inflammatory cells on cerebrospinal fluid (CSF) in an immunosuppressed patient can lead to further investigations, which unravel the presence of cryptococcal meningitis. The aim of this retrospective study was to analyze the spectrum of CSF findings of immunosuppressed patients who were diagnosed to have cryptococcal meningitis. Retrospective analysis of CSF cytospin slides exhibiting pleocytosis and belonging to immunocompromised patients was performed, and these cases were found to have cryptococcal meningitis. Out of 932 cases of CSF (January 2016-July 2017), 10 had pleocytosis and 5 of these 10 cases demonstrated spores of Cryptococcus. Male-to-female ratio was 1:1.5. All the patients were immunocompromised and had CSF leukocytosis. Lymphocytes and monocytes were present in all samples while only one case showed plasma cells. Spores of Cryptococcus were also noted in all the cases and highlighted on India Ink Preparation and Gomori Methenamine Silver stain. All the cases were positive for agglutination-based cryptococcal antigen assay, except one where the test was not done. All immunosuppressed patients having pleocytosis in CSF were found to have cryptococcal infection. Therefore, pleocytosis in CSF in any immunosuppressed patient should raise the suspicion of cryptococcal meningitis. The pathologist and the clinician need to be vigilant in such scenarios to rule out any opportunistic infection and investigate the patient thoroughly for any underlying immunosuppression.

Protection of mice against experimental cryptococcosis using glucan particle-based vaccines containing novel recombinant antigens

Meningitis due to Cryptococcus neoformans is responsible for upwards of 180,000 deaths worldwide annually, mostly in immunocompromised individuals. Currently there are no licensed fungal vaccines, and even with anti-fungal drug treatment, cryptococcal meningitis is often fatal. Our lab previously demonstrated vaccination with recombinant cryptococcal proteins delivered in glucan particles (GPs) protects mice against an otherwise lethal infection. The aim of the present study was to discover additional cryptococcal antigens affording vaccine-mediated protection. Sixteen proteins, each with evidence of extracellularity, were selected for in vivo testing based on their abundance in protective alkaline extracts of an acapsular C. neoformans strain, their known immunogenicity, and/or their high transcript level during human infection. Candidate antigens were recombinantly expressed in E. coli, purified and loaded into GPs. BALB/c and C57BL/6 mice received three subcutaneous injections of GP-based vaccine, and survival was assessed for 84 days following a lethal orotracheal challenge with strain KN99. As with our six published GP-vaccines, we saw differences in overall protection between mouse strains such that BALB/c mice typically demonstrated better survival than C57BL/6 mice. From these studies, we identified seven new proteins which, when administered as GP-vaccines, protect BALB/c and/or C57BL/6 mice against cryptococcal infection. With these results, we expand the pool of novel protective antigens to eleven proteins and demonstrate the potential for selection of highly transcribed extracellular proteins as vaccine targets. These screens highlight the efficacy of GP-subunit vaccines and identify promising antigens for further testing in anti-cryptococcal, multi-epitope vaccine formulations.

Cryptococcal Antigenemia in Human Immunodeficiency Virus Antiretroviral Therapy-Experienced Ugandans With Virologic Failure.

BackgroundDetectable serum or plasma cryptococcal antigen (CrAg) precedes symptomatic cryptococcal meningitis. The World Health Organization recommends CrAg screening for human immunodeficiency virus–positive persons with CD4 count <100 cells/μL initiating antiretroviral therapy (ART). However, an increasing proportion of patients with cryptococcosis are now ART experienced. Whether CrAg screening is cost-effective in those with virologic failure is unknown.MethodsWe retrospectively performed nationwide plasma CrAg testing among ART-experienced Ugandan adults with virologic failure (≥1000 copies/mL) using leftover plasma after viral load testing during September 2017–January 2018. For those who were CrAg positive, we obtained ART history, meningitis occurrence, and 6-month survival via medical records review.ResultsAmong 1186 subjects with virologic failure, 35 (3.0%) were CrAg positive with median ART duration of 41 months (interquartile range, 10–84 months). Among 25 subjects with 6-month outcomes, 16 (64%) survived, 7 (28%) died, and 2 (8%) were lost. One survivor had suffered cryptococcal meningitis 2 years prior. Two others developed cryptococcal meningitis and survived. Five survivors were known to have received fluconazole. Thus, meningitis-free survival at 6 months was 61% (14/23). Overall, 91% (32/35) of CrAg-positive persons had viral load ≥5000 copies/mL compared with 64% (735/1151) of CrAg-negative persons (odds ratio, 6.0 [95% confidence interval, 1.8–19.8]; P = .001). CrAg prevalence was 4.2% (32/768) among those with viral loads ≥5000 copies/mL and 0.7% (3/419) among those with viral loads <5000 copies/mL.ConclusionsIn addition to the CD4 threshold of <100 cells/μL, reflexive CrAg screening should be considered in persons failing ART in Uganda with viral loads ≥5000 copies/mL.

1410. Serious Cryptococcal Infections with Ruxolitinib Use: A Case of Meningitis and a Review of the Literature

BackgroundRuxolitinib is an inhibitor of Janus kinase (JAK) 1 and 2 and is approved for the treatment of myelofibrosis and polycythemia vera. Infectious complications associated with its use include reactivation of herpes simplex, zoster, hepatitis B, and tuberculosis, mucormycosis, and progressive multifocal leukoencephalopathy.MethodsSeven cases of ruxolitinib-associated cryptococcal infections have been reported: three cases of meningitis, two cases of pulmonary disease, and two cases of disseminated disease (Table 1).ResultsWe present a 72-year-old male with a history of JAK-2-positive polycythemia vera with secondary myelofibrosis, and concurrent multiple myeloma who presented with 3 weeks of chronic cough and 3 days of fever with severe bifrontal headache after remodeling a large birdcage in his backyard. The patient was on ruxolitinib, ixazomib, and weekly dexamethasone. Cerebrospinal fluid (CSF) analysis showed an elevated opening pressure of 29 cm of CSF, 173 leucocytes with a predominance of lymphocytes, protein of 87 mg/dL and a normal glucose level. BioFire FilmArray® Meningitis/Encephalitis panel detected targets for Cryptococcus in the CSF, and CSF cryptococcal antigen was positive at 1:4. CSF fungal cultures were subsequently positive for Cryptococcus neoformans, susceptible to liposomal amphotericin B (LAMB) and fluconazole. Ruxolitinib was discontinued, treatment with LAMB and flucytosine (5-FC) was associated with significant improvement of headache over the next week. Repeat CSF analysis in 2 weeks was culture negative with a negative cryptococcal antigen test. The patient completed 3 weeks of LAMB and 5-FC and then transitioned to oral fluconazole for a year.ConclusionRuloxitinib and other JAK inhibitors suppress host immunity through impaired T-cell activation and downregulation of cytokines. The JAK-Signal Transducer and Activator of Transcription pathway is one of the most active pathways in host defense against cryptococcus, and use of JAK inhibitors like ruxolitinib may predispose to severe cryptococcal infections. Bird exposure is a risk factor, making environmental contact counseling for patients on ruxolitinib important. Fluconazole prophylaxis needs to be considered in select cases. Disclosures All authors: No reported disclosures.

1821. Evaluation of Cerebrospinal Fluid White Blood Cell Count Criteria for Use of the BioFire® FilmArray® Meningitis/Encephalitis Panel in Immunocompetent Patients

BackgroundIn 2016, our academic medical center implemented the BioFire® FilmArray® Meningitis/Encephalitis Panel (MEP), which detects 14 viral, bacterial, and fungal pathogens. Institutional guidelines recommended the test be used in nonimmunocompromised patients age ≥2 years only if the cerebrospinal fluid (CSF) white blood cell (WBC) count was >10 cells/mm3.MethodsWe reviewed all MEP performed at our institution over 2 years (January 1, 2017 to December 31, 2018). We collected CSF WBC count, protein, and glucose; MEP results; CSF culture results; and demographics. We excluded children age <2 years, immunocompromised patients, those without a CSF WBC count, and duplicate tests during the same illness.ResultsOf 453 patients, 311 met inclusion criteria. The median age was 51, 51% male. Median CSF indices: WBC/mm3 = 4, protein = 57 mg/dL, glucose = 66 mg/dL. MEP positivity rate = 12% (37/311): viruses (29/37), bacteria (7/37), and fungi (1/37). Positive bacterial/fungal MEP results compared with CSF culture are summarized in Table 1. No clinically significant discordant negative MEP results occurred compared with CSF culture, cryptococcal antigen, or other viral PCR testing. Of the 311 patients, 184 (59%) had ≤10 CSF WBC/mm3. Of these, 4 had positive MEP results: 1 enterovirus, 1 human herpes virus 6 (HHV-6) and 2 varicella zoster virus (VZV). The HHV-6 was judged clinically insignificant. The 2 VZV cases had concomitant shingles and were already on acyclovir. No clinically significant MEP results occurred in 110/311 (35%) patients with ≤ 2 CSF WBC/mm3.ConclusionIn nonimmunocompromised patients, age ≥ 2, with ≤ 10 CSF WBC/mm3 on lumbar puncture, positive MEP results were rare and the clinical significance of the 4 positives was debatable. A hard-stop restriction in this setting could have reduced overall use by up to 59% and resulted in significant cost savings. Lower CSF WBC/mm3 cut-offs could be considered and still improve MEP utilization. Disclosures All Authors: No reported Disclosures.

375. Cryptococcal Antigenemia in Advanced HIV Infection

BackgroundDiagnostic importance of asymptomatic cryptococcal antigenemia is being increasingly recognized in the last few years. Recently, WHO (World Health Organization) has recommended routine screening of CrAg (cryptococcal antigen) among PLHA with CD4 ≤100/mm3, albeit this procedure is not yet adopted by many developing countries including India.MethodsWe conducted a prospective observational study in a large tertiary care center of North India, upon ethical clearance. Latex agglutination test was performed to assess serum CrAg levels, followed by the lumbar puncture for detection of CrAg levels in the CSF. We analyzed the prevalence and treatment outcomes of cryptococcal antigenemia among PLHA with CD4 ≤ 100 cells/mm3. Detailed clinical examination was conducted, with follow-up of upto 3 months. Multivariate analysis was performed for the estimation of risk factors.ResultsThe mean age (years) and BMI (kg/m2) of all the participants were 41.4 ± 11.2 and 22.1 ± 2.6, respectively. Notably, the mean CD4 count (cu.mm) at the time of recruitment was 62.3 ± 20.5. Noteworthy, 62 (60.8%) of the patients were ART naïve. We found 9.8% (n = 10) of the patients were positive for serum CrAg, and only 2.9% (n = 3) had clinical features of meningitis and 6.8% (n = 7) were asymptomatic (subclinical) CrAg positive. Strikingly, 3.9% (n = 4) of the asymptomatic cryptococcal antigenemia patients were also positive for CrAg in CSF, with 1.9% (n = 2) were only serum CrAg positive, and 1 patient was lost to follow-up (Graph 1). Multivariate analysis revealed that patients with long duration of HIV (P = 0.04), headache symptoms (P = 0.004) and possessing features of meningismus (P value=0.08) are more likely to be CrAg positive. Conversely, patients on fluconazole were protective against cryptococcal antigenemia (P = 0.1) as shown in Table 1. Overall mortality observed was 11.3% among advanced HIV patients. Moreover, mortality in CrAg-positive patients was 33.3% in comparison to CrAg-negative patients who had 9% (P = 0.06) in 3-months follow-up.ConclusionCryptococcal antigenemia is common (9.8%) among patients with CD4 count ≤100/mm3 in India. Screening for CrAg should be made routine for PLHA with CD4 count ≤100/mm3 and if required preemptive treatment to be given in this regard. Disclosures All authors: No reported disclosures.

253. Evaluation of a Cryptococcal Antigen Lateral Flow Assay and the Burden of Cryptococcal Disease: A Cohort Study at Grady Memorial Hospital in Atlanta, Georgia

BackgroundWhile Cryptococcus neoformans is a major cause of morbidity and mortality among HIV-infected persons worldwide, there is scarce recent data on disease prevalence in the United States, including in Southeastern states, where HIV rates are high. We sought to determine the prevalence of cryptococcal disease and compare the performance of a cryptococcal antigen (CrAg) lateral flow assay (LFA) vs. latex agglutination (LA) test.MethodsAll patients from Grady Memorial Hospital in Atlanta, Georgia who had a serum or cerebrospinal fluid (CSF) sample sent for CrAg LA testing as part of routine management from November 2017 to July 2018 were included. The LFA was performed on all samples by research staff; results were not available to clinicians. Rates of disease and agreement between the LA test and LFA were calculated.ResultsAmong 467 patients, 570 LA tests were performed; 417 on serum and 153 on CSF (87 patients with multiple tests performed). Mean age was 44 years, and most were male (n = 322, 69%). Most patients had HIV (n = 371, 79%); median CD4 count was 73 cells/mm3 and 77% were not receiving ART. Among HIV-infected individuals, testing was performed equally in the inpatient and outpatient setting. Cryptococcal testing was done in 53 persons without apparent risk factors. Thirty-three (7%) patients had a positive serum or CSF test. Five (1%) patients had both a positive serum and CSF LA test and LFA. While the overall agreement between the LA test and LFA was substantial to high for CSF (κ = 0.71) and serum (κ = 0.93), respectively, there were important discrepancies. Four patients with a negative serum LA test had a positive serum LFA. Five patients had false-positive CSF LA tests, determined by negative CSF LFA testing, India ink, and CSF and fungal cultures. All were treated with amphotericin and flucytosine with one patient experiencing a severe anaphylactic reaction to amphotericin.ConclusionWe found a moderately high rate of cryptococcal disease and important discrepancies between the LA test and LFA. The LFA appeared to be more sensitive for cryptococcemia and more specific for meningitis. Clinical implications of these findings include earlier detection and treatment of cryptococcemia, and averting unnecessary treatment of meningitis with costly medications associated with high rates of adverse events. Disclosures All authors: No reported disclosures.

1713. Impact of False-Positive Low-Titer Cryptococcal Antigen Testing

BackgroundAt University of Kentucky (UK) HealthCare, the transition from latex agglutination testing (Remel, Lenexa, KS) to IMMY Cryptococcal Antigen Lateral Flow Assay (CrAg LFA) occurred in September 2016. A few months later, it was noticed that several cryptococcal cases were diagnosed with weak positive test results where the diagnosis could not be confirmed by additional testing. The purpose of this study was to analyze the characteristics of these patients, and to assess the interventions they received based on positive results.MethodsThis was a retrospective study of the patients with positive CrAg LFA treated at UK HealthCare from November to December 2016. Low antigen titers (≤ 1:20) were considered to be false positive if repeat testing with the Remel Cryptococcal Latex assay, IMMY latex and IMMY microwell EIA were negative, cultures and histopathology were negative and there was no clear clinical evidence of infection.ResultsDuring this 2-month period, CrAg LFA was positive in 12 patients. The diagnosis of cryptococcosis could not be confirmed by additional testing in 9 (75%) individuals. Cirrhosis/liver disease was present in 3 (33.3%) patients, 5 (55.6%) underwent lumber puncture and antifungal therapy was administered in 8 (88.9%) patients (Table 1). CrAg LFA was false positive in 1/2 (50%) HIV, 3/3 (100%) transplant, and 5/7 (71.4%) non-HIV/non-transplant patients (Figure 1). Among the false positives, 4 (44.4%) patients had titer of 1:5, two (22.2%) had 1:20, and the original positive screen was not detected upon titration in 3 (33.3%) other patients. One HIV patient received a complete treatment course for unconfirmed cryptococcal meningitis because an LP could not be performed.ConclusionFalse-positive low CrAg LFA titers led to unnecessary tests, antifungal treatments and prolonged hospitalization in some patients. One-third of these individuals had cirrhosis/liver disease. Other institutions also reported false-positive low CrAg LFA titers. As a result, the company staged a recall of the specific lot and corrected the problem in reagent manufacturing. Low-positive titers using CrAg LFA should be interpreted carefully and further testing should be considered as determined by the clinical situation. Disclosures All authors: No reported disclosures.