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Results for “STAINS AND STAINING”

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PubMed2001

Indocyanine green selectively stains the internal limiting membrane.

PURPOSE: To demonstrate whether indocyanine green stains the inner limiting membrane of the retina or residual vitreous cortex. METHODS: We report on the intraoperative staining patterns of the vitreomacular interface in 10 eyes of 10 consecutive patients who underwent vitrectomy with indocyanine green staining for macular hole formation and diffuse diabetic macular edema. RESULTS: In five eyes of five patients with macular holes, indocyanine green staining of the macula after posterior vitreous detachment resulted in an immediate visibility of a discernable membrane that was not previously seen. In five eyes of five patients with diffuse diabetic macular edema and adherent cortical vitreous, indocyanine green failed to stain the vitreomacular interface. After peeling off the residual vitreous cortex, however, a discernable membrane could be identified using indocyanine green dye again. Light and transmission electron microscopy revealed the inner limiting membrane as the membrane that had been stained and removed in all specimens. CONCLUSION: Indocyanine green selectively stains the inner limiting membrane. Staining of the vitreomacular interface using indocyanine green as a vital dye enables the surgeon to distinguish between the residual vitreous cortex and the inner limiting membrane, and it allows safer and easier removal of the inner limiting membrane.

Basement Membrane↗

PubMed1992

Cryptosporidium parvum sporozoite staining by propidium iodide.

Modified Ziehl-Neelsen (ZN) acid-fast stain is the usual method for detection of Cryptosporidium oocysts in feces. Propidium iodide permitted us to stain free or intra-oocyst sporozoites. With the ZN method only 3-5% of the oocysts purified from three human and one experimentally infected lamb dichromate-preserved feces were stained by carbol fuchsin. These fuchsin-stained oocysts were free of intact sporozoites as identified by propidium iodide staining. Treatment with 10% formalin or 0.5% sodium hypochlorite increased the percentage of acid-fast stained oocysts and thus the sensitivity of acid-fast staining. Treatment with sodium hypochlorite induced intra-oocyst sporozoite alterations as demonstrated by flow cytometric analysis of the oocysts' DNA content. Propidium iodide staining of fixed oocysts is a simple and rapid method to visualize sporozoites and to assess oocyst preservation after different treatments.

Animals↗

PubMedDate not supplied

Does precipitation reduce tissue staining by indocyanine green dye solutions?

PURPOSE: Indocyanine green (ICG) dye precipitates when mixed with certain ophthalmic irrigation solutions. The purpose of this study is to investigate whether this precipitation reduces ICG staining of the anterior lens capsule. METHODS: ICG was diluted with each of the following solutions: BSS Plus, physiological saline, or Opeguard Neo. The products were then examined for green precipitate by light microscopy. The tissue staining capability of each ICG solution was tested at two different concentrations (0.5% and 0.0625%) in porcine lenses, regardless of whether the solution contained precipitate. RESULTS: Green precipitate was observed in both concentrations of ICG solutions diluted with BSS Plus, but not in the solutions diluted with either physiological saline or Opeguard Neo. As assessed with light microscopy, staining of the anterior lens capsule appeared weaker for all 0.0625% ICG solutions compared to the corresponding 0.5% ICG solutions. The precipitate that formed in the 0.5% ICG solution diluted with BSS Plus had little effect on the staining quality of the anterior lens capsule. In contrast, the 0.0625% ICG solution diluted with BSS Plus (w/precipitate) showed weaker staining in the lens capsule compared to the other two 0.0625% ICG solutions (w/o precipitate). CONCLUSION: These results suggest that precipitation of ICG may weaken its capability to stain the anterior lens capsule or other transparent ocular tissues. Therefore, ICG solutions that do not form a precipitate may be more capable of staining tissues at lower concentrations. As for other possibilities to explain the deterioration in staining, the effect of the composition of BSS Plus should also be considered.

Animals↗

PubMed2003

A simple method for immunocytochemical staining with multiple rabbit polyclonal antibodies.

We report a single step, simple, repeatable, rapid and reliable technique for simultaneous immunocytochemical staining with two or more rabbit polyclonal antibodies. This technique, which we have dubbed the "Pretty Poly" method, is based on conjugating the antibodies with commercially available, fluorophore-tagged Staphylococcal protein-A (SP-A). Staining is illustrated at the calyx type presynaptic nerve terminal of the chick ciliary ganglion with antibodies directed against three nerve terminal proteins: neurofilaments of the axonal cytoskeleton, and two secretory vesicle proteins, SV2 and cysteine string protein (CSP). Images were deblurred with an iterative deconvolution protocol. Staining with a single polyclonal antibody was bright and had a resolution approaching light microscope limit. Treatment with two different polyclonal antibodies conjugated with contrasting dye-tagged protein-A resulted in double staining without significant crossover that was fully equivalent to the standard primary/secondary technique. The same single step protocol was used to stain with all three rabbit polyclonal antibodies or to combine the technique with a standard monoclonal primary/secondary antibody stain. Thus, the Pretty Poly protocol is a highly flexible, simple and yet effective staining technique that essentially solves the problem of co-staining with multiple polyclonal rabbit antibodies.

Animals↗

PubMed1989

Comparison of stains for image segmentation and measurement of nuclear parameters by computerised image analysis using IBAS 2000.

Nuclear measurements using image analysis largely depend upon the quality of the image presented for digitization. To investigate the effects of nuclear stains on image segmentation of nuclei, serial sections of kidney were stained by eleven different methods and presented to an IBAS 2000 interactive image analysis system (Kontron Bildanalyse) via a Zeiss IIIRS microscope at x 800 magnification and a Siemens K30 video camera. Digitised grey level images of each field were processed by an interactive technique and by an automatic segmentation procedure (thresholding). Nuclear areas were measured by each method and the results compared. We conclude that of the stains assessed the uncounterstained haematoxylins offer the best image segmentation for nuclear measurements. Thresholding techniques are suitable for performing measurements using these stains, particularly when additional interactive techniques are used to reject unwanted structures and to separate overlapping nuclei after segmentation. Comparable areas stained with five of the stains were studied to see if the staining techniques themselves affected nuclear area. Our results show that the use of different stains will substantially affect measurements of nuclear dimensions.

Cell Nucleus↗

PubMed1999

TUNEL staining of inner ear structures may reflect autolysis, not apoptosis.

A recent study (Usami et al., 1997) using the TUNEL method has suggested that age-related cell death in the senescence-accelerated mouse inner ear is due to apoptosis. TUNEL staining detects not only apoptosis but also late necrosis or autolysis because it detects DNA breaks. Autolysis may occur in inner ear structures during fixation. To determine whether or not age-related cell death is due to apoptosis, TUNEL staining of the inner ear of normal mice should be understood. However, studies of TUNEL staining of the normal inner ear have not yet been reported. We investigated whether the fixation method or the interval between the death of normal mice and the initiation of fixation influences the results of TUNEL staining of the inner ear. Marginal cells of the stria vascularis and hair cells of the saccule were TUNEL-positive, irrespective of the fixation method or the interval between death and fixation. Interdental cells, Reissner membrane cells, fibrocytes in the suprastrial region, and inner and outer hair cells were also occasionally stained. Transmission electron microscopy showed no morphological characteristics of apoptosis in the hair cells of the saccule. Moreover, patterns of TUNEL staining in the normal and senescence-accelerated mouse inner ear were similar. These stained tissues may require a high level of oxygen, making them more susceptible to autolysis. We concluded that the results of TUNEL staining in the inner ear require confirmation by morphological studies.

Animals↗

PubMed2003

Staining methods applied to glycol methacrylate embedded tissue sections.

The use of glycol methacrylate (GMA) avoids some technical artifacts, which are usually observed in paraffin-embedded sections, providing good morphological resolution. On the other hand, weak staining have been mentioned during the use of different methods in plastic sections. In the present study, changes in the histological staining procedures have been assayed during the use of staining and histochemical methods in different GMA-embedded tissues. Samples of tongue, submandibular and sublingual glands, cartilage, portions of respiratory tract and nervous ganglion were fixed in 4% formaldehyde and embedded in glycol methacrylate. The sections of tongue and nervous ganglion were stained by H&E. Picrosirius, Toluidine Blue and Sudan Black B methods were applied, respectively, for identification of collagen fibers in submandibular gland, sulfated glycosaminoglycans in cartilage (metachromasia) and myelin lipids in nervous ganglion. Periodic Acid-Schiff (PAS) method was used for detection of glycoconjugates in submandibular gland and cartilage while AB/PAS combined methods were applied for detection of mucins in the respiratory tract. In addition, a combination of Alcian Blue (AB) and Picrosirius methods was also assayed in the sublingual gland sections. The GMA-embedded tissue sections showed an optimal morphological integrity and were favorable to the staining methods employed in the present study. In the sections of tongue and nervous ganglion, a good contrast of basophilic and acidophilic structures was obtained by H&E. An intense eosinophilia was observed either in the striated muscle fibers or in the myelin sheaths in which the lipids were preserved and revealed by Sudan Black B. In the cartilage matrix, a strong metachromasia was revealed by Toluidine Blue in the negatively-charged glycosaminoglycans. In the chondrocytes, glycogen granules were intensely positive to PAS method. Extracellular glycoproteins were also PAS positive in the basal membrane and in the region occupied by the lamina externa and reticular fibers surrounding each smooth muscle cells of the blood vessels. In the epithelial cells of the respiratory tract, acid and neutral mucins were histochemically detected by AB and PAS methods, respectively. Moreover, granules containing acid and neutral mucins were revealed in purple by AB and PAS concomitantly. In the sublingual gland sections, a distinct affinity of acid mucins by AB (in turquoise-blue) and collagen fibers by Picrosirius (in red) was obtained when these methods were combined. Although some routine dyes used in paraffin sections have showed a weak stain in historesin sections, our results showed that different dyes could be applied in GMA sections if modified staining procedures were assayed. Therefore, appropriate staining contrast and, thus, detection of one or different substances in a same section can be acquired in association to the good morphological resolution provided by GMA.

Alcian Blue↗

PubMed1997

Systematic investigations of the contrast results of histochemical stainings of neurons and glial cells in the human brain by means of image analysis.

The investigation of neurohistological specimens by image analysis has become an important tool in morphological neuroscience. The problems which arise during the processing of these images are non-trivial, especially if a pattern recognition of cells in the imaged tissue is intended. One of the major problems faced concerns the segmentation of structures of interest, whether cells or other histologic structures. The segmentation problem is often the result of an inappropriate staining procedure. For serious image analysis to be performed, the material under investigation must be optimally prepared. Spatially complex patterns, e.g. fuzzy-like neighbouring neurons, are easy to recognize for humans. But the integrative and associative performance of current artificial neuronal network schemes is too low to achieve the same recognition quality as humans do. Therefore, a general analysis of staining characteristics was performed, especially with respect to those stains which are relevant to object segmentation. Although most image analytical investigations of tissues are based on stained samples, a study of this type has not been previously conducted. Of the stains and procedures evaluated, the gallocyanin chrome alum combination staining provided the best stain contrast. Furthermore, this staining method shows sufficient constancy within different parts of the human brain. Even the fine nuclear textures are differentiable and can be used for further pattern recognition procedures.

Adult↗

PubMed2004

Combined histochemical staining, RNA amplification, regional, and single cell cDNA analysis within the hippocampus.

The use of five histochemical stains (cresyl violet, thionin, hematoxylin & eosin, silver stain, and acridine orange) was evaluated in combination with an expression profiling paradigm that included regional and single cell analyses within the hippocampus of post-mortem human brains and adult mice. Adjacent serial sections of human and mouse hippocampus were labeled by histochemistry or neurofilament immunocytochemistry. These tissue sections were used as starting material for regional and single cell microdissection followed by a newly developed RNA amplification procedure (terminal continuation (TC) RNA amplification) and subsequent hybridization to custom-designed cDNA arrays. Results indicated equivalent levels of global hybridization signal intensity and relative expression levels for individual genes for hippocampi stained by cresyl violet, thionin, and hematoxylin & eosin, and neurofilament immunocytochemistry. Moreover, no significant differences existed between the Nissl stains and neurofilament immunocytochemistry for individual CA1 neurons obtained via laser capture microdissection. In contrast, a marked decrement was observed in adjacent hippocampal sections stained for silver stain and acridine orange, both at the level of the regional dissection and at the CA1 neuron population level. Observations made on the cDNA array platform were validated by real-time qPCR using primers directed against beta-actin and glyceraldehyde-3 phosphate dehydrogenase. Thus, this report demonstrated the utility of using specific Nissl stains, but not stains that bind RNA species directly, in both human and mouse brain tissues at the regional and cellular level for state-of-the-art molecular fingerprinting studies.

Aged↗

PubMed2002

Recovery of intact proteins from silver stained gels.

Silver stained proteins of a wide molecular weight (MW) range (20-116 kDa) were successfully recovered by both electroblot and electroelution. The recovery was demonstrated for nanogram loads of proteins separated by SDS-PAGE and visualized by silver staining methods compatible and incompatible with mass spectrometry (MS). It was shown that the alcohol/acid and glutaraldehyde fixation steps present in a number of staining procedures did not prevent recovery of intact proteins from gels. It was found that the recovery of intact proteins from silver stained gels was substantially increased upon pre-equilibration in a buffer containing the reducing agent, dithiothreitol (DTT). The effect of destaining on the recovery of silver stained proteins was also investigated. Comparable recovery of intact proteins within a wide MW range from silver stained gels with and without destaining step was demonstrated. Recovery of model proteins from gels visualized using silver staining method compatible with MS showed 52 to 76% yield of that from the unstained gel, depending upon method of the transfer. Comparison of the recovery of intact proteins from gels visualized using other staining procedures was also made. The above findings have implications as to the supposed irreversible nature of protein "fixation" inside polyacrylamide matrix, and confirm lack of binding of proteins in the gel to metal silver deposited on its surface. This method has the potential to be suitable for direct characterization of proteins by matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) without additional purification steps.

Electrophoresis, Polyacrylamide Gel↗

PubMed1998

Rapid staining of ultrathin sections with the use of a microwave oven.

The microwave oven has many potential applications, ranging from tissue fixation to staining for light and electron microscopy. This study was planned to speed up the staining of ultrathin sections. The first set of grids was stained conventionally with uranyl acetate and Reynold's lead citrate solutions. The other grids were stained with the same solutions by microwave irradiation. The electron micrographs of grids stained using the microwave technique were as satisfactory as the grids stained conventionally. Microwave-treated grids demonstrated more uniform staining and less precipitate. The use of a microwave oven shortened staining time by approximately 38 min.

Animals↗

PubMed2001

Influence of sample preparation, staining procedure and analysis conditions on bull sperm head morphometry using the morphology analyser integrated visual optical system.

The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200,000-300,000 spermatozoa/microl. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+). Diff-Quik (DIF), haematoxylin (HEM). Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40x or 100x oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF-stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40x objective yielded optimal results concerning sperm recognition and digitization. The 100x objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000-500 000 spermatozoa/microl). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000-300 000 spermatozoa/microl appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria).

Animals↗

PubMed2001

Capsular collagen staining of follicular thyroid neoplasms by picrosirius red: role in differential diagnosis.

A key criterion in the diagnosis of thyroid follicular carcinoma is capsular invasion, but invasion cannot always be demonstrated histologically. Since invasion is likely to evoke reactions in the capsular collagen, we examined the effects of invasion on capsular collagen with the picrosirius orange-red (PSR) staining technique for collagen. Under polarized light, the color of PSR-stained collagen varies as a function of the structural and biochemical properties of the collagen fibers. Capsules of widely invasive carcinomas (n = 10), minimally invasive carcinomas (n = 10), and adenomas (n = 28) were stained with the PSR method. Carcinomas were assessed along the thickened capsule for sites of definite invasion, minimal invasion, and no evidence of invasion. In adenomas, sites of thickened capsules (similar to carcinomas) were compared to sites of thin capsules. All foci were evaluated for the color and color intensity of collagen fibers. We found a significantly higher frequency of yellow-green collagen fibers than of orange-red fibers at sites of invasion, whereas orange-red fibers significantly predominated at non-invaded sites. In a minority of cases both colors occurred but the non-dominant color was of lesser intensity in all but 1 case. There were no significant differences in staining between minimally and widely invasive carcinomas. Thick capsules of adenomas consistently stained with an intense orange-red color, although weakly stained yellow-green fibers were also observed in some of these cases. We conclude that PSR staining can provide diagnostically useful information in capsular samples of carcinomas, when both color and color intensity of PSR staining are evaluated at the same site. Specifically, intense yellow-green birefringence of collagen in a thickened capsule is additional evidence for capsular invasion.

Adenoma↗

PubMed2000

Assessment of different methods for staining Helicobacter pylori in endoscopic gastric biopsies.

The recent implication of Helicobacter pylori in the pathogenesis of gastritis-peptic ulcer syndrome and its relevance for the development of upper gastrointestinal malignancy warrant efficient methods for the detection and demonstration of the organism in biopsy specimens. We have compared 5 staining methods, namely, haematoxylin and eosin (H & E), immunohistochemistry (IHC), the silver staining HpSS, the alcian yellow-toluidine blue (Leung) method (A-Y) and Genta staining, for the demonstration of the organism in gastric biopsies taken from antrum, body and fundus of 118 patients who presented to our hospital with upper gastrointestinal symptoms. We found no significant differences in the efficacy of H & E, IHC, HpSS and A-Y in the demonstration of H. pylori in all 3 gastric sites. The least reproducible stain in our hands was the Genta stain. We conclude that H & E is adequate for the initial assessment of gastric biopsies in symptomatic upper gastrointestinal patients. This is because it is a well-tested, cheap and easy staining method, requiring a relatively short period of time to perform, with highly reproducible results. It has an added advantage of enabling simultaneous assessment of morphological changes accompanying H. pylori infection. When the density of the organism is expected to be low, we recommend addition of HpSS staining because of its high sensitivity and low cost. The disadvantages of the other staining methods (IHC, A-Y and Genta) are discussed.

Adolescent↗

PubMed1955

Electron densitometry of stained virus particles.

Methods are described for determining the relative mass of particles in electron microscope specimens through the measurement of photographic densities in recorded images. These methods were applied to a quantitative study of the amounts of electron stains that could be associated with the particles of tomato bushy stunt virus (BSV) and tobacco mosaic virus (TMV). In the pH range above 2 where the viruses are stable, the amount of stain absorbed is too small to produce adequate contrast in the electron microscope. Maximum stain absorption was achieved at pH about 1 where with several reagents and combinations of reagents the mass of BSV could be increased to about four times that of the unstained particles. Optimum results were obtained with phosphotungstic acid alone or in combination with Pt, Th, or La ions. Since the pH conditions for high stain absorption are normally destructive, morphology is satisfactorily preserved only when the phosphotungstic acid is applied in concentrations of 10 per cent or greater or when the use of destructive reagents is preceded by a preliminary fixation under mild conditions. Maximum staining of TMV increased the mass of the particles to about two times that of the unstained. Estimates of the mass of heavily stained BSV particles indicate that their density is 3.3 gm./cm.(3) The high internal hydration of BSV probably accounts for the greater stain absorption and penetration compared to those of TMV which has very low or zero internal hydration. Anomalous images resulting from the use of electron stains are shown and discussed.

Coloring Agents↗

PubMed1994

The relative sensitivity of special stains and culture in open lung biopsies.

The author compared the efficacy of Gram's, methenamine silver, and acid-fast stains on tissue sections to conventional culture methods in vitro for detecting infection in 110 open lung biopsies (99 inflammatory, 11 neoplastic). Twenty-one cases of clinically significant infection were found (19%). Gram's stain and culture had sensitivities of 100% and 80%, respectively. However, 21 of the cultures grew clinically irrelevant organisms and 2 Gram's stains originally were misinterpreted. In contrast, methenamine silver and acid--fast stains were significantly more sensitive (80% and 100%) than their corresponding cultures (20% and 50%) and no methenamine silver or acid-fast stains were misinterpreted. In addition, methenamine silver stain had a higher positive predictive value than culture (100% vs. 60%). In conclusion, both methenamine silver and acid-fast stains are more sensitive than culture for detecting significant pathogens from open lung biopsies. Gram's stain is as sensitive as bacterial culture but is often misinterpreted. Moreover, interpretation of bacterial cultures is complicated by the frequent growth of clinically insignificant organisms.

Adult↗

PubMed1996

Immunocytochemical staining of cytologic specimens. How helpful is it?

The contribution of immunocytochemical (ICC) staining to the diagnosis of a range of cytologic specimens reported over a 20-month period was retrospectively assessed. A total of 194 cases (1.6% of total workload) were stained for diagnostic purposes in this period. Immunocytochemical staining was determined to be helpful in 75.8% of cases. In body fluid cases (105), staining was most often performed to assist with the discrimination of mesothelial cells and metastatic malignancy. Immunocytochemical staining was helpful in 82% of fluid specimens, with the preliminary diagnosis confirmed in 64%, refined in 8%, and revised in 10% of cases. In fine-needle aspiration (FNA) specimens, staining was helpful in 69%, resulting in a refinement of diagnosis in 55%, and confirming the preliminary diagnosis in 14%. The preliminary diagnosis was revised in a single FNA case. Immunocytochemical staining was particularly valuable in assisting the subclassification of poorly differentiated malignancies. A more precise diagnosis was possible following consideration of ICC results in 68% of cases with a preliminary diagnosis of poorly differentiated malignancy. Immunocytochemical staining was also helpful in identifying the primary site of metastatic carcinoma in six FNA cases. It is concluded that the selective use of well-chosen panels of antibodies can be very helpful in resolving diagnostic difficulties in cytologic specimens. In particular, ICC may be an invaluable aid to the diagnosis of difficult serous effusion specimens and in the sub-typing of poorly differentiated malignancy.

Antibodies↗

PubMed2000

Special histologic stains are rarely beneficial for the evaluation of HIV-related gastrointestinal infections.

During a 28-month period, endoscopic mucosal biopsy specimens from all HIV-infected patients were submitted for routine histologic evaluation. Immunoperoxidase staining for cytomegalovirus and herpesvirus antigens (esophagus), mycobacterial and fungal staining, and Gram staining of mucosal biopsy specimens were done. Special fungal and acid-fast stains were selectively performed in patients with absolute CD4 cell counts of less than 200 cells per microliter (200 x 10(6)/L) and/or with diarrhea and or wasting syndrome. Treatment was based on the endoscopic and histologic findings, and long-term follow-up was performed. The 121 symptomatic HIV-infected patients underwent 221 upper and/or lower endoscopies with 285 biopsy sites. The sensitivity and specificity of H&E staining for the diagnosis of gastrointestinal cytomegalovirus were 97% and 100%, respectively. The results of fungal and mycobacterial stains neither altered therapy nor identified previously undiagnosed infections in any patient. Long-term follow-up revealed no patient in whom an infection was missed on routine H&E, which affected outcome. Routine H&E staining is accurate for the diagnosis of gastrointestinal opportunistic infections in HIV-infected patients. Special histologic stains for fungal, mycobacterial, and viral infections did not increase the diagnostic yield or alter medical therapy but doubled the costs.

AIDS-Related Opportunistic Infections↗